Effect of oridonin on invasion and migration of human lung cancer NCI-H460 cells

AIM：To investigate the effect of oridonin on the invasion and migration of human lung cancer NCI-H460 cells. METHODS：NCI-H460 cells were divided into high-dose (HD), middle-dose (MD) and low-dose (LD) oridonin groups (cultured with 40, 20 and 10 μmol/L of oridonin, respectively, as experimental groups), and normal (N) group (treated without oridonin as control). The cell growth was observed. The cell proliferation was detected by MTT assay. Boyden chamber was used to determine the cell invasive capacity. The cell migration was also measured. The levels of MMP-2 and MMP-9 were assayed by Western blotting. RESULTS：The cell counts in the experimental groups were lower than that in N group. The cell proliferation was inhibited as the inhibitory rates were 48.94%, 36.17% and 19.15% for HD group, MD group and LD group, respectively. The numbers of the invasive cells were 26.67±5.16 for HD group, 36.17±5.08 for MD group, and 44.33±5.50 for LD group. The migration rates in the experimental groups were lower than that in N group. The expression of MMP-2 and MMP-9 decreased dependent on the oridonin dose as follows: HD group < MD group < LD group < N group. CONCLUSION：Oridonin inhibits the invasion and migration of NCI-H460 lung cancer cells, and reduces the expression of MMP-2 and MMP-9.     

Long non-coding RNA TTTY15 expression in osteosarcoma and its effect on viability and invasion ability of osteosarcoma cells

AIM:To observe the expression of long noncoding RNA TTTY15 in osteosarcoma tissues and cell lines and to explore its effect on the viability and invasion ability of osteosarcoma cell lines. METHODS:qPCR was used to detect the expression of TTTY15 in 11 cases of osteosarcoma and its adjacent tissues. The mRNA levels of TTTY15 in osteosarcoma cell lines (143B, Saos2, MG-63, U2OS and HOS) and human osteoblast cell line hFOB1.19 were also tested. TTTY15 was down-regulated after transfected with small interfering RNA in MG-63 cells, the cell line with the highest level of TTTY15. The effect of TTTY15 knockdown on the viability of MG-63 cells was measured by CCK-8 assay. The cell cycle distribution was analyzed by flow cytometry. The effect of TTTY15 knockdown on the cell invasion ability was detected by Transwell assay. The levels of miR-216b-5p and FOXM1 mRNA were detected by qPCR, and the changes of the related proteins were determined by Western blot. RESULTS:Compared with the adjacent tissues, the expression of TTTY15 increased in the osteosarcoma tissues (P<0.01). Compared with the human osteoblast cell line, the expression of TTTY15 increased in the osteosarcoma cell lines (P<0.05), and the level of TTTY15 in the MG-63 cells was the highest (P<0.01). After knockdown of TTTY15 expression in the MG-63 cells, the cell viability was decreased (P<0.05), cell cycle progression was inhibited (P<0.01), and the cell invasion ability was decreased (P<0.01). The expression of miR-216b-5p was increased (P<0.01) and the expression of FOXM1 mRNA was decreased (P<0.01). The protein expression of FOXM1, CDK4, cyclin D1, MMP-2 and N-cadherin was decreased, while the protein expression of E-cadherin was increased (P<0.05). CONCLUSION:The expression of TTTY15 is increased in the osteosarcoma tissues and cell lines. The low expression of TTTY15 inhibits the cell viability and invasion ability of osteosarcoma cells. The possible mechanism is that the knockdown of TTTY15 expression results in the increase in miR-216b-5p expression and the down-regulation of FOXM1 expression.     

Long non-coding RNA PVT1 regulates effect of miR-551 on migration and invasion abilities of ovarian cancer through Wnt signaling pathway

AIM: To investigate the expression of long non-coding RNA PVT1 in ovarian cancer and the role of PVT1 in migration and invasion abilities of ovarian cancer cells.METHODS: The expression of PVT1 in ovarian cancer tissue, normal ovarian tissue and different ovarian cancer cell lines was detected by qPCR. Transwell assay was used to detect the invasion ability of ovarian cancer cells after PVT1 silencing. The migration ability of the ovarian cancer cells after PVT1 silencing was detected by scratch test. The interaction between PVT1 and microRNA (miR)-551 was analyzed by dual-luciferase reporter assay. The effect of miR-551-inhibitor on the invasion and migration abilities of ovarian cancer cells after PVT1 silencing was detected by Transwell assay and scratch test. The expression of Wnt signaling pathway-related proteins was determined by Western blot after PVT1 silencing. The effects of PVT1 silencing on tumor weight and volume of ovarian cancer were examined by subcutaneous tumor transplantation in nude mice.RESULTS: The expression of PVT1 in ovarian cancer tissue was significantly higher than that in normal ovarian tissue (P<0.05). The expression level of PVT1 in ovarian cancer cell line ES-2 was the highest. PVT1 silencing inhibited the invasion and migration abilities of the ovarian cancer cells. After PVT1 silencing, miR-551-inhibitor promoted the invasion and migration abilities of the ovarian cancer cells. The expression of Wnt signaling pathway-related proteins was decreased after PVT1 silencing (P<0.05). Compared with negative control group, the tumor volume and weight in PVT1-siRNA group were significantly decreased (P<0.05).CONCLUSION: PVT1 plays an important role in the development of ovarian cancer. PVT1 regulates the invasion and migration abilities of ovarian cancer cells through Wnt signaling pathway.     

Effect of long non-coding RNA-671 on proliferation and invasion of esophageal squamous-cell carcinoma cells

AIM: To detect the expression of long non-coding RNA-671 (lnc671) in esophageal squamous-cell carcinoma cell lines and to investigate the effect of lnc671 on the malignant phenotype of esophageal squamous-cell carcinoma cells. METHODS: The level of lnc671 in the esophageal squamous-cell carcinoma cells was detected by RT-qPCR. Specific lnc671 small interfering RNA (siRNA) used to explore the effects of lnc671 on proliferation, colony formation, invasion and migration abilities of esophageal squamous-cell carcinoma cells. RESULTS: The database of GEPIA analysis showed that increased expression of lnc671 was associated with shorter survival in the patients of esophageal cancer (P<0.05). Compared with normal immortalized esophageal epithelial cells, lnc671 was highly expressed in a variety of esophageal squamous-cell carcinoma cell lines. lnc671 knock-down significantly inhibited the growth, colony formation ability, migration and invasion abilities of esophageal squamous-cell carcinoma cells(P<0.01). CONCLUSION: The expression of lnc671 is increased in various esophageal squamous-cell carcinoma cell lines. Knock-down of lnc671 expression inhibits the malignant phenotype of esophageal squamous-cell carcinoma cells.     

Study on expression of long non-coding RNA in colon cancer tissues and adjacent tissues

AIM: To screen the differentially expressed long non-coding RNA (lncRNA) in colon cancer, and to explore its expression in colon cancer tissues and adjacent tissues. METHODS: The "Colon adenocarcinoma:Person neoplasm cancer status" which consisted of 36 cases of colon cancer tissues and 29 cases of normal colonic tissues was downloaded from the lncRNAtor database. The candidate genes were selected from these differentially expressed lncRNAs based on artificial criterion (P<0.01; fold change ≥ 2 or<0.5) and then validated by real-time PCR in 60 pairs of colon cancer tissues and adjacent tissues. RESULTS: A total of 50 lncRNAs were differentially expressed in colon cancer tissues, including 28 up-regulated and 22 down-regulated (P<0.01). The verifying results displayed that HNF1A-AS1 and ZDHHC8P1 were up-regulated (P<0.01), and SUZ12P expression was down-regulated (P<0.05), but the expression of AC069513.3 was not statistically significant between colon cancer tissues and adjacent tissues. The abilities of HNF1A-AS1, ZDHHC8P1, SUZ12P and AC069513.3 to discriminate the colon cancer from normal adjacent tissue by the ROC curve with an AUC of 0.729 (sensitivity 78%, specificity 67%), 0.617 (sensitivity 68%, specificity 55%), 0.689 (sensitivity 66%, specificity 55%) and 0.518 (sensitivity 52%, specificity 48%) were observed. CONCLUSION: Long non-coding RNA HNF1A-AS1 and ZDHHC8P1 are up-regulated and SUZ12P is down-regulated in colon cancer tissues, suggesting that they may be involved in the pathogenesis of colon cancer.     

Effect of CD97 gene silencing by small interfering RNA on migration and invasion of gastric carcinoma cell lines

AIM: To investigate the effect of CD97 gene silencing by small interfering RNA(siRNA) on migration and invasion of gastric carcinoma cell lines. METHODS: Gastric carcinoma cell lines AGS and MGC803 were used in the study. Four pairs of siRNA were designed according to the sequence of CD97 gene and synthesized chemically. The siRNAs were transfected into the gastric carcinoma cell lines. Forty-eight hours after transfection, the total RNA was extracted and the mRNA expression of CD97 was detected by real-time RT-PCR so as to screen the most effective siRNA. The protein level of CD97 was also measured by fluorescence-activated cell sorting (FACS) 72 h after Transfection. The abilities of migration and invasion were evaluated by Transwell test. The viability of the cells was measured by MTT method. RESULTS: Real-time RT-PCR and FACS revealed that CD97-siRNA notably down-regulated CD97 expression at both mRNA and protein levels. The mRNA level decreased by (89.34±9.95)% and (95.42±1.93)% in AGS and MGC803 cells,respectively. The protein levels of CD97^EGF and CD97^stalk in AGS cells decreased by (19.29±3.45)% and (30.11±5.93)%,respectively. The protein levels of CD97^EGF and CD97^stalk in MGC803 cells decreased by (26.25±5.73)% and (16.22±3.23)%,respectively. No change of the cell viability after siRNA transfection was observed. The cell number of migration and invasion in AGS cells was decreased by (67.63±12.03)% and (68.02±15.63)%,respectively. The cell number of migration and invasion in MGC803 cells was decreased by (14.92±2.03)% and (22.09±5.43)%,respectively. CONCLUSION: The siRNA effectively inhibits CD97 expression and restrains the migration and invasion capacities of gastric carcinoma cell lines, suggesting that CD97 plays an important role in the metastasis of gastric cancer.     

CASC2/miR-18a/BTG3 signaling inhibits cell migration and invasion of non-small cell lung cancer

AIM: To explore the function and molecular mechanism of long non-coding RNA CASC2 in non-small-cell lung cancer (NSCLC) cell migration and invasion. METHODS: RT-qPCR and Western blot were used to detect the expression of CASC2, microRNA-18a (miR-18a) and BTG3 in human bronchial epithelial cell line 16-HBE, and NSCLC cell lines A549 and H1299. The interaction between CASC2 and miR-18a or miR-18a and BTG3 was predicted by bioinformatics software and verified by double-luciferase reporter assays. Transwell assays were performed to detect the migration and invasion abilities of the NSCLC cells. RT-qPCR and Western blot were used to determine the regulatory effects of CASC2 on miR-18a and BTG3 expression. RESULTS: Compared with 16-HBE cells, the expression of CASC2 and BTG3 was significantly down-regulated in the NASCL cells, while miR-18a was significantly over-expressed (P<0.05). CASC2 acted as a molecular sponge for miR-18a, and BTG3 was verified to be a target gene of miR-18a. Up-regulation of CASC2 inhibited the migration and invasion abilities of NSCLC cells, while exogenous restoration of miR-18a stimulated cell migration and invasion abilities. In addition, exogenously over-expressed miR-181a reversed the promoting effect of CASC2 on BTG3 protein expression. CONCLUSION: CASC2 promotes BTG3 expression by negatively regulating miR-18a, and then inhibits the migration and invasion abilities of NSCLC cells.     

Effects of curcumin on migration and invasion of myeloma cells

AIM：To explore the effect of curcumin on the migratory and invasive abilities of myeloma cells. METHODS：shRNA expression plasmid was transfected into RPMI8226 cells to knock down IQ motif-containing GTPase-activating protein 1 (IQGAP1). The expression of IQGAP1 in RPMI8226 cells tranfected with shIQGAP1 or shRNA negative control, and in un-transfected RPMI8226 cells was detected by Western blotting. All the cells in RPMI8226-shIQGAP1 group, RPMI8226-shRNA negative control group and un-transfected RPMI8226 group were treated with curcumin at various concentrations. The migratory and invasive abilities of the RPMI8226 cells were measured by Transwell chamber and Matrigel assays. The expression of IQGAP1 at mRNA and protein levels in the RPMI8226 cells treated with curcumin was also determined by RT-PCR and Western blotting. RESULTS：The expression of IQGAP1 decreased when IQGAP1 gene was knocked down by shRNA. The migration and Matrigel invasion tests showed that the number of cells moving into under chamber of Transwell decreased in RPMI8226-shIQGAP1 group in comparison with the other 2 groups. Curcumin decreased the migratory and invasive abilities of RPMI8226-shRNA negative control cells and un-transfected RPMI8226 cells, which was not related to the curcumin concentratory. The migratory and invasive abilities of RPMI8226-shIQGAP1 cells showed no significant difference when treated with curcumin at various concentrations. The expression of IQGAP1 at mRNA and protein levels decreased in the RPMI8226 cells treated with curcumin. CONCLUSION：Curcumin decreases the migratory and invasive abilities of myeloma cells via inhibition of IQGAP1 expression.     

Gastrin promotes the migration and invasion of gastric cancer cells

AIM：To study the effects of gastrin on the migration and invasion of gastric cancer cells in vitro. METHODS：The migration and invasion of gastric cancer AGS and SGC-7901 cells after treated with gastrin at concentrations of 10 nmol/L and 100 nmol/L were studied by wound-healing assay and Transwell migration and invasion assay. The cell proliferation was analyzed by MTT colorimetric method. The concentration of matrix metalloproteinase 2 (MMP-2) in the culture medium was detected by ELISA. The AGS and SGC-7901 cells without treating with gastrin served as control cells. RESULTS：Compared with the control cells, the migration and invasion of AGS cells and SGC-7901 cells were significantly increased after treated with gastrin at concentrations of 10 nmol/L and 100 nmol/L. In control, 10 nmol/L gastrin and 100 nmol/L gastrin groups, the mean numbers of the migrating cells were 56.0, 88.1 and 106.4/view in AGS cells and 52.8, 91.0 and 113.3/view in SGC-7901 cells, and the mean numbers of the invasive cells were 78.4, 118.7 and 141.6/view in AGS cells and 87.3, 124.6 and 147.4/view in SGC-7901 cells, respectively. The numbers of the migrating cells and invasive cells in 100 nmol/L gastrin group were higher than those in 10 nmol/L gastrin group. The cell proliferation rate and the concentration of MMP-2 in the culture medium in gastrin treatment groups were higher than those in control group. CONCLUSION：Gastrin promotes the migration and invasion of gastric cancer cells in a dose-dependent manner by increasing the MMP-2 secretion, which may be the key mechanism in the proliferation, invasion and metastasis of the cancer cells in vivo.     

Aldolase A level in malignant pleural effusion and its effects on proliferation,migration and invasion of human lung adenocarcinoma cells

AIM：
To investigate the levels of aldolase A (ALDOA), carcinoembryonic antigen (CEA) and lactate dehydrogenase (LDH) in malignant pleural effusion (MPE) from patients with lung cancer and tuberculous pleural effusion (TBPE) from patients with tuberculous pleurisy, and to explore the effects of ALDOA on the proliferation, migration and invasion of human lung adenocarcinoma A549 cells. METHODS：Pleural effusion samples including 65 cases of MPE and 35 cases of TBPE were collected, and the levels of ALDOA, CEA and LDH were detected by ELISA and chemiluminescence assay. After A549 cells were treated with different concentrations of ALDOA, the proliferation, migration and invasion of the cells were investigated by MTT assay, scratch test, Matrigel assay and Transwell invasion assay. RESULTS：The levels of ALDOA, CEA and LDH in MPE were (46.8±21.4) μg/L, (82.2±56.6) μg/L and (755.8±382.5) U/L, respectively, which were significantly higher than those in TBPE ［(23.9±17.2) μg/L, (12.6±9.7) μg/L and (388.4±163.9) U/L, respectively; P<0.01］. The concentration of ALDOA in MPE from adenocarcinoma patients ［(71.7±32.1) μg/L］ was significantly higher than that in MPE from squamous-cell carcinoma patients ［(21.3±14.6) μg/L, P<0.05］. The concentrations of ALDOA in MPE and TBPE were positively correlated with the concentrations of CEA and LDH (P<0.01 or P<0.05). ALDOA enhanced the proliferation, migration and invasion of A549 cells in a concentration-dependent manner. CONCLUSION：The expression level of ALDOA in MPE is significantly higher than that in TBPE, especially in MPE from lung adenocarcinoma patients. There are highly positive correlations between ALDOA and CEA, ALDOA and LDH in pleural effusion. ALDOA concentration-dependently promotes the proliferation, migration and invasion of A549 cells.     

Effect of norcantharidin on proliferation and invasion of human breast cancer cell line SKBR3 in vitro

AIM: To investigate the effect of norcantharidin（NCTD）on proliferation and invasion of human breast cancer cell line SKBR3 in vitro and its anticancer mechanisms.METHODS: MTT assay was used to determine SKBR3 cell proliferation. Light and FACScan were used to detect apoptosis and cell cycle. The invasiveness of SKBR3 was evaluated by the adhesion test，Matrigel experiment and the crossing-river test.RESULTS: NCTD had inhibitive effects on growth of SKBR3 cells in a dose and time-dependent manner, with the IC₅₀ value of 12.5 mg/L at 24 h．The cells treated with 10 mg/L NCTD for 24 h and 48 h showed typical apoptotic morphology and hypodiploid peak before G₁ phase. The cell cycle was arrested at G₂/M phase. The apoptosis percentage was up to 3.44% and 6.17%, and the G₂/M percentage was up to 35.82% and 38.70%. NCTD also could inhibit obviously the adhesion, movement and invasive capability simulating human basement membrane of SKBR3. Its effect was also in a dose-dependent manner. In the NCTD-treated group, crossing-river time was prolonged significantly and passing-membrane cells markedly decreased. CONCLUSION: NCTD in vitro inhibits not only the proliferation and growth of human breast cancer cells but also invasion and metastasis of the cells at relatively low concentration. NCTD shows prominent anti-tumor effects.     

Inhibitory effect of anti-miR-196a on invasion and migration of human pancreatic cancer PANC-1 cells

AIM:To investigate the expression of miR-196a in different pancreatic cancer cell lines and to observe the effect of anti-miR-196a on the biological behaviors of human pancreatic cancer PANC-1 cells. METHODS:The expression of miR-196a in the pancreatic cancer cells was examined by real-time quantitative PCR. Anti-miR-196a was chemically synthesized and transfected into PANC-1 cells by Lipofectamine 2000. The cell proliferation was measured by CCK-8 assay. The apoptosis was determined by flow cytometry. Matrigel invasion assay was performed to examine the migration and invasion of the tumor cells. The wild-type and mutant-type NF-κB inhibitor α(NFKBIA) 3'UTR luciferase reporter vectors were constructed. The relative activity of Renilla luciferase was detected to confirm the binding site of miR-196a on NFKBIA mRNA. RESULTS:The expression of miR-196a in human pancreatic cancer cell lines was significantly higher than that in human pancreatic ductal epithelial H6c7 cells. The expression of anti-miR-196a in miR-196a group was down-regulated. After transfected with miR-196a, no change of cell proliferation and apoptosis was observed, but the abilities of invasion and migration were reduced(P<0.01). Compared with negative control, wild-type NFKBIA3'UTR or mutant-type NFKBIA 3'UTR, cotransfection of anti-miR-196a and wild-type NFKBIA 3'UTR significantly increased the relative activity of Renilla luciferase. CONCLUSION:miR-196a is one of the oncomiRs and may be a target microRNA of human pancreatic cancer for gene therapy.     

Effect of hypoxia on invasion and migration of lung carcinoma cells and its molecular basis

AIM: To investigate the effect of hypoxia on the invasion and migration of lung carcinoma cells. METHODS: Lung carcinoma cell H128 was exposed to normoxia (air, 5% CO₂), hypoxia (5% O₂,5% CO₂,90% N₂) or anoxia (95% N₂,5% CO₂) conditions for 48 hours. The migration ability of the cells was assayed by wound healing methods. The invasiveness ability was determined with HABM-HEM model. The cells exposed to hypoxia were inoculated under skin in nude mice, and then the growth of the tumor and the rate of metastasis to lymph node or lung were observed. The expression of E-cadherin and β1-integrin on the cells were also assayed by flow cytometry. RESULTS: Compared with normoxic group, the invasiveness and migration in hypoxic group were increased. The rates of tumorgenesis and metastasis to lung in anoxic group were evidently decreased. The expression of E-cadherin was decreased, and the expression of β1-integrin was increased in hypoxic group. In anoxia group, the invasiveness, migration, the expression of E-cadherin and β1-integrin were all decreased. CONCLUSIONS: Moderate hypoxia down-regulated the expression of E-cadherin, up-regulated the expression of β1-integrin, and increased the invasiveness and metastasis of carcinoma cells. Serious hypoxia decreased the expression of adhesive molecules, and the proliferation, invasiveness and migration were also decreased.     

Sinomenine inhibits viability,migration and invasion of human ovarian cancer SKOV3 cells

AIM:To investigate the effect of sinomenine on the viability, migration and invasion of human ovarian cancer SKOV3 cells and its possible mechanism. METHODS:The SKOV3 cells were treated with sinomenine at different concentrations for 12 h, 24 h and 48 h. CCK-8 assay was employed to detect the effects of sinomenine on the viability of the SKOV3 cells. Flow cytometry was used to analyze the cell cycle distribution. The cell migration and invasion abilities were measured by Transwell assay. Western blot was used to determine the protein levels of cyclin A, cyclin D1, E-cadherin and matrix metalloproteinase-9 (MMP-9). RESULTS:Sinomenine remarkably inhibited the viability of SKOV3 cells and IOSE80 cells in a time-dependent and dose-dependent manner (P<0.05), and the IC₅₀ values of 48 h were 2.12 mmol/L and 17.35 mmol/L, respectively. In a dose-dependent manner, sinomenine induced G₀/G₁ and S phase arrest in SKOV3 cells (P<0.05), suppressed the migration and invasion abilities of SKOV3 cells (P<0.05), down-regulated the protein levels of cyclin A, cyclin D1 and MMP-9 (P<0.05), and up-regulated the protein level of E-cadherin (P<0.05). CONCLUSION:Sinomenine inhibits the viability, migration and invasion of human ovarian cancer SKOV3 cells most likely via down-regulation of the protein levels of cyclin A, cyclin D1 and MMP-9, and up-regulation of the protein level of E-cadherin.     

Effects of p21-activated kinase 6 on invasion and migration of human non-small-cell lung cancer A549 cells

AIM：To investigate the effects of p21-activated kinase 6 (PAK6) on the invasive and migratory abilities of human non-small-cell lung cancer A549 cells. METHODS：The expression of PAK6 mRNA in A549 cells, human bronchial epithelial (HBE) cells, non-small-cell lung cancer tissues and paired adjacent non-tumor tissues was measured by real-time PCR. After A549 cells were transfected with siRNA-PAK6 (siPAK6) or negative control (NC) for 48 h, the expression of PAK6 at mRNA and protein levels was measured by real-time PCR and Western blotting, respectively. The invasion and migration of A549 cells were detected by Matrigel invasion assay and Transwell migration assay. The cytoskeletal changes were observed with FITC-phalloidin staining under confocal microscope. RESULTS：The level of PAK6 mRNA in A549 cells was higher than that in HBE cells (3.50±1.16 vs 1.12±0.42, P<0.05). The level of PAK6 mRNA in non-small-cell lung cancer tissues was higher than that in paired adjacent non-tumor tissues (5.13±1.33 vs 1.08±0.37, P<0.05). The expression of PAK6 protein decreased by 72% in A549 cells transfected with siPAK6 (P<0.05), and the level of PAK6 mRNA significantly decreased in A549 cells transfected with siPAK6 (3.72±0.75 vs 0.69±0.21, P<0.05). Matrigel invasion assay and Transwell migration assay demonstrated that knockdown of PAK6 markedly attenuated the invasion and migration of A549 cells (P<0.05). The cytoskeletal actin remodeling and reduction of stress fibers in A549 cells transfected with siPAK6 were observed under confocal microscope. CONCLUSION：PAK6 may affect the invasive and migratory abilities of non-small-cell lung cancer cells by cytoskeletal actin remodeling.     

Long non-coding RNA HOTAIR modulates proliferation and apoptosis of acute lymphoblastic leukemia cells

AIM: To explore the influence of long non-coding RNA HOTAIR on the proliferation and apoptosis of acute lymphoblastic leukemia cells via the regulation of glucocorticoid receptor (GR).METHODS: The expression le-vels of HOTAIR and GR mRNA in human bone marrow stromal cell line HS-5 and human acute lymphoblastic leukemia cell lines MOLT-4, CCRF-CEM and CEM-C1 were examined by RT-qPCR. HOTAIR was knocked down by siRNA in acute lymphoblastic leukemia cells. CCK-8 assay was used to assess the cell viability, and the effect of si-HOTAIR on the proli-feration of CEM-C1 cells was evaluated by BrdU method. The effect of si-HOTAIR on apoptosis of CEM-C1 cells was examined by Hoechst 33342 staining and Caspase-Glo^® 3/7 assay. Western blot was utilized to examine the protein level of GR.RESULTS: The expression level of HOTAIR in acute lymphoblastic leukemia cells was significantly increased as compared with normal human bone marrow stromal cells (P<0.01). The viability and proliferation of acute lymphoblastic cells was inhibited, the apoptosis was induced, and the anti-proliferation effect of dexamethasone on CEM-C1 cells was enhanced after knockdown of HOTAIR expression (P<0.01). The expression of GR was up-regulated at both mRNA and protein levels (P<0.01).CONCLUSION: Long non-coding RNA HOTAIR may modulate the viability, proliferation and apoptosis of acute lymphoblastic leukemia cells via a GR regulatory way.     

Growth inhibition of colorectal cancer HCT116 cells by lincRNA-p21 through STAT3 signaling pathway

AIM: To study the effect of long intergenic non-coding RNA-p21 (lincRNA-p21) on the growth inhibition of colorectal cancer HCT116 cells via STAT3 signaling pathway. METHODS: The human colorectal cancer cell line HCT116 was used to construct the cells with over-expression of lincRNA-p21 by transfection of pcDNA-lincRNA-p21, and negative control cells were also set up. After transfection, the expression level of lincRNA-p21 was detected by RT-qPCR. The cell viability and proliferation were examined by MTT assay and plate colony formation assay, respectively. The protein levels of STAT3 and phosphorylated STAT3 (p-STAT3) were determined by Western blot. After STAT3 signaling pathway activator SD19 was used to treat the colorectal cancer HCT116 cells with over-expression of lincRNA-p21, Western blot was used to detect the protein levels of STAT3 and p-STAT3, MTT assay was used to measure the viability of the cells, and flow cytometry analysis was used to determine the cell apoptosis. RESULTS: Compared with control group and pcDNA group, the expression of lincRNA-p21 in pcDNA-lincRNA-p21 group was significantly up-regulated, the cell proliferation was inhibited, and the protein levels of STAT3 and p-STAT3 were significantly decreased (P<0.05). After treatment with STAT3 activator SD19, the protein levels of STAT3 and p-STAT3 in pcDNA-lincRNA-p21+SD19 group were higher than those in pcDNA-lincRNA-p21 group, the cell viability was increased, and the apoptotic rate was decreased significantly (P<0.05). CONCLUSION: Over-expression of lincRNA-p21 inhibits the growth of colorectal cancer HCT116 cells. STAT3 signaling pathway activator abolishes the growth inhibitory effect of lincRNA-p21 over-expression. lincRNA-p21 inhibits the growth of colorectal cancer cells by inhibiting the activation of STAT3 signaling.     

Effects of siRNA-mediated nestin silencing on invasion and migration of human esophageal cancer ECA109 cells

AIM: To investigate the effect of small interference RNA(siRNA)-mediated silencing of nestin gene on the invasion and migration of human esophageal cancer ECA109 cells and the possible mechanism. METHODS: The esophageal cell line ECA109 was transfected with siRNA targeting nestin and the cell invasion and migration abilities were observed. The expression of nestin, MMP2, MMP9, VEGF, and total and nuclear β-catenin proteins in the transfec-ted cells were determined by real-time PCR and Western blot. RESULTS: Compared with control group, the expression of nestin at mRNA and protein levels was significantly down-regulated in the ECA109 cells transfected with nestin-siRNA, so was the expression of MMP2, MMP9, VEGF, and total and nuclear β-catenin proteins. The levels of invasion and migration capacities of ECA109 cells transfected with nestin-siRNA were lower than those in the cells transfected with control-siRNA. CONCLUSION: Knockdown of nestin expression significantly inhibits the invasion and migration of the esophageal cancer cells, which may act via suppressing β-catenin translocation to the nucleus and influencing the expression of MMP2, MMP9 and VEGF.     

Effect of formononetin on viability,migration and invasion of ovarian cancer cells in vitro

AIM To observe the effect of formononetin on the viability, migration and invasion of ovarian cancer cells, and to explore its mechanism. METHODS Human ovarian serous cystadenocarcinoma SKOV-3 cells were cultured in vitro. The cells were treated with formononetin at 0, 25, 50 and 100 μmol/L for 48 h. The cell viability was measured by MTS assay. The migration and invasion abilities of the SKOV-3 cells were detected by scratch wound assay and Transwell assay. RT-qPCR and Western blot were used to detect the mRNA and protein levels of E-cadherin and matrix metalloproteinase-9 (MMP-9). RESULTS The viability of SKOV-3 cells was decreased with the increase in the formononetin concentration compared with control group (P<0.01). The wound migration distance of the cells in 50 μmol/L formononetin group was less than that in control group (P<0.01). The number of invasive SKOV-3 cells across the Transwell sub-compartment was significantly decreased in 50 μmol/L formononetin group compared with control group (P<0.01). The mRNA and protein levels of E-cadherin in 50 μmol/L formononetin group were significantly higher than those in control group (P<0.01), while the mRNA and protein levels of MMP-9 in 50 μmol/L formononetin group were significantly lower than those in control group (P<0.01). CONCLUSION Formononetin inhibits the migration and invasion abilities of ovarian cancer SKOV-3 cells by increasing expression of E-cadherin and decreasing expression of MMP-9.