Pirfenidone inhibits TGF-β1-induced differentiation of myofibroblast in human lung fibroblast populations

ATM: To investigate the effect of pirfenidone on transforming growth factor-β1 (TGF-β1)-induced fibroblast-to-myofibroblast transition in vitro. METHODS: The cell viability was measured by MTT assay. The proliferation of human lung fibroblasts (HLFs) was detected by EdU incorporation. Migratory and invasive abilities were measured by Boyden chamber assay. The α-smooth muscle actin (α-SMA) protein expression was determined by Western blot and immunofluorescence. The mRNA expression of α-SMA and type Ⅰ and Ⅲ collagens was evaluated by RT-qPCR. RESULTS: Pirfenidone at different concentrations (0.1, 0.2, 0.3, 0.5 and 0.8 mg/L) had no cytotoxic effect on the HLFs, and pirfenidone at 0.2 mg/L was used for the intervention. Pretreatment of the HLFs with 0.2 mg/L pirfenidone prior to TGF-β1 not only markedly suppressed the changes of proliferation, migration, invasion and reorganization of actin cytoskeleton in the HLFs (P<0.01﹚,but also down-regulated the expression of α-SMA and type C and Ⅲ collagens triggered by TGF-β1 ﹙P<0.05﹚.CONCLUSION: Pirfenidone has an inhibitory effect on TGF-β1-induced activated cell functions and fibroblast-to-myofibroblast transition in HLFs.     

NBP promotes angiogenesis of HUVECs by activating VEGF/VEGFR2-Notch1/Dll4 signal pathway

AIM: To investigate the effects of DL-3-n-butylphthalidle (NBP) on angiogenesis of human umbilical vein endothelial cells (HUVECs) and the role of vascular endothelial growth factor (VEGF)/VEGF receptor 2(VEGFR2)-Notch1/Delta-like ligand 4 (Dll4) signaling pathway in this process. METHODS: The serum-free medium and anoxic tank were used to simulate the conditions of hypoxia and ischemia (H/I). HUVECs were divided into control group, H/I group, H/I+NBP_high group and H/I+NBP_low group. The HUVECs in control group were conventionally cultured, and those in H/I group were cultured under H/I intervention. The HUVECs in H/I+NBP_high group were treated with NBP at 20 μmol/L under H/I intervention. The HUVECs in H/I+NBP_low group were treated with NBP at 5 μmol/L under H/I intervention. The cell viability of each group was measured by CCK-8 assay. The migration ability of the HUVECs in each group was detected by cell scratch test. The vessel formation ability of the HUVECs was examined by in vitro angiogenesis assay. The expression of VEGFR2, Notch1 and Dll4 at mRNA and protein levels was determined by qPCR and Western blot, and the expression of VEGF was determined by qPCR and ELISA. RESULTS: NBP increased the viability of HUVECs, and promoted the migration ability and the formation of blood vessels in vitro under H/I intervention. These effects of NBP at high dose were more significant than those at low dose. NBP increased the expression of VEGF, VEGFR2, Notch1 and Dll4 at mRNA and protein levels (P<0.05). CONCLUSION: NBP promotes HUVECs to form blood vessels under H/I intervention. The mechanism may be related to the activation of VEGF/VEGFR2-Notch1/Dll4 signaling pathway.     

Mechanism of Sos2 inhibiting NFATc1-mediated podocyte injury in diabetic nephropathy

AIM: To observe the effect of high glucose (HG) stimulation on the expression of guanine nucleotide exchange factor Sos2 (Son of Sevenless homolog 2) in mouse podocytes, and to explore the role of Sos2 in HG-induced podocyte damage and its possible molecular mechanisms. METHODS: The expression of Sos2 in the podocytes of diabetic nephropathy patients was observed by immunofluorescence staining and laser confocal microscopy. In vitro, the Sos2 expression at mRNA and protein levels in immortalized podocytes with HG (30 mmol/L glucose) stimulation for 48 h was determined by the methods of RT-PCR, Western blot and immunofluorescence. Using Western blot, immunofluorescence and wound-healing assay, the expression of podocin, the translocation of NFATc1 into the nucleus and the podocyte migration with or without Sos2 silencing or overexpression were analyzed. The expression of downstream target genes for NFATc1 was detected by RT-PCR. RESULTS: The expression of Sos2 was significantly decreased in the podocytes of diabetic nephropathy patients and in vitro cultured podocytes with HG stimulation (P<0.05). When Sos2 was silenced, the expression of podocin was significantly decreased, the migration ability of podocytes was increased, and the translocation of NFATc1 into the nucleus was increased (P<0.05). In contrast, after overexpression of Sos2 in the podocytes with HG stimulation, the podocin expression level was obviously higher, and the podocyte migration ability and the translocation of NFATc1 into the nucleus were decreased (P<0.05).CONCLUSION: Sos2 may attenuate the diabetic nephropathy-induced podocyte injury by inhibiting NFATc1.     

Tumor necrosis factor-related apoptosis-inducing ligand participates in injury of vascular endothelial cells induced by rapamycin in vitro

AIM: To investigate the effects of rapamycin on apoptosis, proliferation, migration ability and tumor related apoptosis inducing ligand(TRAIL) in cultured human umbilical vein endothelial cells(HUVECs). METHODS: Cultured HUVECs were treated with rapamycin at the concentrations of 0, 1, 10 and 100 μg/L for 24 h. The cell proliferation was measured by CCK-8 method. The cell migration ability was detected by Transwell chambers and wound healing test. The apoptotic index of HUVECs was quantitatively determined by measuring the activation of caspase-3. The morphological changes of the apoptotic cells were observed by DAPI staining. The expression of TRAIL was detected by Western blotting. RESULTS: A 24 h-incubation with rapamycin(1-100 μg/L) caused significant cell loss associated with the increase in apoptosis, as quantified by the determination of caspase-3 activity(P<0.01) in HUVECs. Obvious apoptotic morphology was observed by DAPI staining in HUVECs incubated with rapamycin. Rapamycin at the concentrations of 1-100 μg/L also impaired the migration ability of HUVECs(P<0.01). In addition, rapamycin(10-100 μg/L) inhibited the proliferation of HUVECs, whereas rapamycin at 1 μg/L had no such effect(P<0.01). Rapamycin(10-100 μg/L) also induced TRAIL expression in a dose-dependent manner(P<0.01). CONCLUSION: Rapamycin induces apoptosis, and inhibits the proliferation and migration of HUVECs. The up-regulation of TRAIL might be related to the injury of vascular endothelial cells caused by rapamycin.     

Effect of advanced glycosylation end products on function of human adipose tissue-derived stem cells in vitro

AIM：To explore the effect of advanced glycosylation end products (AGEs) on the function of human adipose-derived stem cells (hADSCs) in promoting wound healing. METHODS：hADSCs were isolated by conventional method in vitro and divided into control bovine serum albumin (BSA) group, low-dose AGE-BSA group and high-dose AGE-BSA group. The proliferation and migration of hADSCs with different treatments were determined by WST-8 assay and Transwell assay, respectively. In addition, the expression of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and insulin-like growth factor-1(IGF-1) at mRNA and protein levels was determined by real-time PCR and ELISA analysis. RESULTS：Compared with control group, the proliferation and migration abilities were significantly inhibited in the hADSCs of AGE-BSA group. The mRNA expression of VEGF, HGF and IGF-1 in AGE-BSA group was obviously lower than that in control group. The contents of VEGF, HGF and IGF-1 in hADSCs-conditioned me-dium in AGE-BSA group were also obviously lower than those in control group. CONCLUSION：AGEs alter the intrinsic properties of hADSCs and impair their functions in promoting wound healing, thus affecting the therapeutic potential of hADSCs in the treatment of diabetic ulcers.     

Homocysteine activates Rho kinase signaling pathway to promote viability and migration of rat basilar arterial vascular smooth muscle cells

AIM:To investigate the regulatory effects of homocysteine (Hcy) on the viability and migration of rat basilar arterial smooth muscle cells (BASMCs) and its potential molecular mechanisms. METHODS:BASMCs were isolated, cultured in vitro and treated with Hcy at different concentrations. The cell viability was measured by CCK-8 assay, and the activation of Rho kinase pathway was measured by Western blot. The cells were treated with Hcy at fixed concentration (1 mmol/L), and ROCK inhibitor Y-27632 was also used. The cell cycle distribution was analyzed by flow cytometry. The cell migration ability was detected by wound healing assay and Transwell assay. The activation of antioxidant enzymes, including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and the level of malondialdehyde (MDA) were measured for determining the status of oxidative stress.RESULTS:Hcy increased the viability of BASMCs and the protein expression of GTP-RhoA and ROCK2 in a dose-dependent manner (P<0.05). Compared with the cells treated with Hcy for 24 h, the cells treated with Hcy for 48 h had enhanced viability (P<0.05). Compared with control group, treatment with Hcy increased cell population in S phase and decreased cell population in G₀/G₁ phase, while pre-incubation with Y-27632 reversed Hcy-induced G₁/S phase transition in BASMCs (P<0.05). The cell migration rate in Hcy treatment group was remarkably higher than that in control group(P<0.05), while pre-incubation with Y-27632 reversed Hcy-induced cell migration (P<0.05). Furthermore, Hcy inhibited the activation of SOD and GSH-Px, accompanied with increased MDA level (P<0.05). Compared with Hcy treatment group, pre-incubation with Y-27632 increased the activation of SOD and GSH-Px, but decreased MDA level (P<0.05). CONCLUSION:Homocysteine induces the viability and migration of rat BASMCs, and its mechanisms may be related to activation of Rho kinase pathway.     

Effects of marrow stromal cell-mediated microenvironment on human lung adenocarcinoma A549 cells

AIM: To investigate the effects of marrow stromal cell line HS-5 on human lung adenocarcinoma A549 cells in the tumor microenvironment. METHODS: The effects of HS-5 cell-conditioned medium (HS-5-CM) on the viability and migration ability of A549 cells were detected by MTT assay and wound-healing assay. After treatment with HS-5-CM, the expression of CX3C chemokine receptor 1 (CX3CR1) at mRNA level in the A549 cells was examined by qPCR. The protein levels of p-ERK and ERK in the A549 cells treated with MAPK/ERK pathway inhibitor U0126 were observed by Western blot, the migration ability of the A549 cells was measured by wound-healing assay, and the protein expression of CX3CR1 was determined by Western blot. RESULTS: HS-5-CM promoted the viability and migration ability of the A549 cells (P<0.01). The expression of CX3CR1 at mRNA level in the A549 cells was increased after treatment with HS-5-CM. MAPK/ERK inhibitor U0126 inhibited the activation of MAPK/ERK signaling pathway (P<0.01), and reduced the migration ability (P<0.01) and the expression of CX3CR1 (P<0.05) in the A549 cells. CONCLUSION: HS-5-CM significantly promotes the A549 cell viability and migration ability. Activation of MAPK/ERK signaling pathway and the expression of CX3CR1 may play a important role in this process.     

Paeonol affects viability and migration ability of hepatocellular carcinoma cells by up-regulating miR-424-3p and inhibiting PI3K/AKT signaling pathway

AIM To investigate the effect of paeonol on the viability and migration ability of hepatocellular carcinoma cells and its molecular mechanism. METHODS Human hepatocellular carcinoma Hep3B cells was treated with paeonol at different concentrations (50, 100, 200 and 400 mg/L). The cell viability was measured by CCK-8 assay to determine the optimal drug concentration. The Hep3B cells were divided into normal control (NC) group, paeonol group, miR-NC group, miR-424-3p group, paeonol+anti-miR-NC and paeonol+anti-miR-424-3p group. The expression level of miR-424-3p was detected by RT-qPCR. The migration ability was detected by Transwell assay. The protein levels of cyclin D1, matrix metalloproteinase 2 (MMP2), MMP9 and PI3K/AKT signaling pathway-related molecules were determined by Western blot. RESULTS Paeonol intervention inhibited the viability of Hep3B cells in a concentration-dependent manner (P<0.05). The concentration of paeonol at 200 mg/L was selected for the following study. Paeonol intervention inhibited the protein expression of MMP2 and MMP9 in the Hep3B cells, and inhibited the migration ability of the Hep3B cells. Paeonol intervention promoted the expression of miR-424-3p in the Hep3B cells (P<0.05). Over-expression of miR-424-3p inhibited the expression of cyclin D1, MMP2 and MMP9 in the Hep3B cells and inhibited cell viability and migration ability (P<0.05). Inhibition of miR-424-3p reversed the effect of paeonol on the viability and migration ability of the Hep3B cells (P<0.05). Paeonol inhibited phosphorylation levels of PI3K and AKT in the Hep3B cells and inhibited the activation of PI3K/AKT signaling pathway (P<0.05). Inhibition of miR-424-3p reversed the effect of paeonol on PI3K/AKT signaling pathway in the Hep3B cells (P<0.05). CONCLUSION Paeonol inhibits the viability and migration ability of hepatocellular carcinoma cells by up-regulating miR-424-3p and inhibiting PI3K/AKT signaling pathway.     

Effects of axitinib on biological behavior of adrenocortical carcinoma cell line SW-13

AIM: To investigate the effects of axitinib on the biological behavior of adrenocortical carcinoma cell line SW-13. METHODS: CCK-8 assay was used to measured the viability of SW-13 cells treated with axitinib at different concentrations. The cell cycle distribution was analyzed by flow cytometry. The apoptotic rate was also analyzed by flow cytometry with Annexin V/PI double staining. Wound healing experiment and Transwell invasion assay were used to observe cell migration and invasion abilities,respectively. The protein levels of vascular endothelial growth factor receptor 2(VEGFR2), extracellular regulated protein kinases 1/2 (ERK1/2) and p-ERK1/2 were determined by Western blot. RESULTS: After treated with axitinib, the viability of SW-13 cells was significantly inhibited, the cell cycle was blocked in G₂/M phase, and the apoptosis rate was increased. The migration and invasion abilities of SW-13 cells were markedly inhibited by axitinib (P<0.01). The protein levels of VEGFR2 and p-ERK1/2 in the SW-13 cells were significantly decreased with axitinib treatment (P<0.01). CONCLUSION: Axitinib inhibits the viability, blocks the cell cycle, promotes cell apoptosis, and inhibits the migration and invasion abilities of SW-13 cells. The mechanism may be related to inhibition of VEGFR2 expression and reduction of ERK1/2 phosphorylation.     

SDF-1α/CXCR4 axis promotes migration and invasion of pancreatic cancer cells through inducing epithelial-mesenchymal transition

AIM:To investigate the role of SDF-1α/CXCR4 axis in pancreatic cancer cell migration and invasion.METHODS:The mRNA expression of CXCR4 in 4 pancreatic cancer cell lines was detected by RT-qPCR. The migration and invasion abilities of PANC-1 cells with the axis activated by exogenous SDF-1α or inhibited by CXCR4 inhibitor AMD3100 were detected by Transwell assays. The cell viability was measured by MTS assay. The protein expression of the epithelial-mesenchymal transition (EMT)-related molecules in the cells treated with exogenous SDF-1α or AMD3100 was determined by Western blot.RESULTS:All of the 4 pancreatic cancer cell lines expressed CXCR4 mRNA, while the PANC-1 cell line expressed the most. Exogenous SDF-1α promoted the migration and invasion abilities of PANC-1 cells, which was inhibited by AMD3100. The PANC-1 cells treated with exogenous SDF-1α for 72 h grew faster, while SDF-1α combined with AMD3100 made little significance to the viability of PANC-1 cells. Exogenous SDF-1α induced EMT of PANC-1 cells by up-regulating the expression of SNAIL and TWIST, and AMD3100 reversed this effect.CONCLUSION:SDF-1α/CXCR4 axis enhances the migration and invasion abilities of pancreatic cancer cells through inducing EMT.     

Fibrinolytic activity and its mechanisms in various leukemic cell lines

AIM:To study the relation between expression of uPAR and annexinⅡ and fibrinolytic activity in various leukemic cell lines. METHODS:The plasma activity was measured under the reaction between cells of NB4, SHI-1, K562, Jurkat, Raji and plaminogen by chromogenic assay. The protein expressions of uPAR and annexinⅡin cells of NB4, SHI-1, K562, Jurkat, Raji were detected by flow cytometry method. The mRNA expressions of uPAR and annexinⅡin cells of NB4, SHI-1, K562, Jurkat, Raji were detected by RT-PCR. RESULTS:The plasma activity in SHI-1 cells and NB4 cells were higher obviously than that in Raji, K562 and Jurkat cells. The protein expression ratio of uPAR and annexinⅡ in NB4 cells were (13.15±1.61)% and (95.97±1.19)%， respectively, they were (99.00±0.26)%, (90.35±2.15)% respectively in SHI-1 cells, and they were lower in K562, Jurkat, Raji cells. The expression of annexinⅡ mRNA in NB4 cells was higher than that in SHI-1 cells, and they were undectectable in K562 and Jurkat cells. The expression of uPAR mRNA in NB4 and SHI-1 cells were higher than that in Jurkat and K562 cells. The expression of uPAR mRNA in Raji cells was undectectable. CONCLUSION:The primary hyperfibrinolysis in leucocythemia cells was observed, and relation was closely with the expression of annexinⅡ. It might be the main reason for bleeding and disseminated intravascular coagulation in patients with acute promyelocytic leukemia and acute monocytic leukemia.     

Effect of HC-1119 on biological behaviors of triple-negative breast cancer BT549 cells

AIM: To explore the effect of new artificially synthesized androgen receptor (AR) antagonist HC-1119 on the biological function of triple-negative breast cancer (TNBC) BT549 cells and the molecular mechanism. METHODS: The AR expression was assessed in different human breast cancer cell lines MDA-MB-231, T47D, MCF-7, SKBR3 and BT549 by Western blot. The TNBC BT549 cells with AR positive expression were treated with HC-1119. The cell viability was measured by CCK-8 assay. The apoptosis rate and cell cycle distribution were analyzed by flow cytometry. The migration and invasion abilities were detected by Transwell assay in vitro. The protein expression of E-cadherin, vimentin and P21 was determined by Western blot. RESULTS: AR was positively expressed in BT549 cells. HC-1119 inhibited the cell viability in a time-and dose-dependent manner (P<0.05), increased the percentage of apoptotic cells and the percentage of S-phase cells significantly, repressed the migration and invasion abilities (P<0.05), and decreased P21 expression at protein level (P<0.01). No influence on the expression of E-cadherin and vimentin in the BT549 cells was observed. CONCLUSION: AR antagonist HC-1119 decreases the viability, migration ability and invasion ability, enhances the apoptosis, and arrests the cell cycle distribution of TNBC BT549 cells. HC-1119 represses the viability of BT549 cells by down-regulating P21 expression, while the process of epithelial-mesenchymal transition is not involved in the inhibition of cell migration.     

Effects of SphK1 and FAK on epithelial-mesenchymal transition in colon cancer HCT116 cells

AIM: To investigate the effects of sphingosine kinase l(SphK1) and focal adhesion kinase(FAK) on the epithelial-mesenchymal transition(EMT) of human colon cancer HCT116 cells. METHODS: Human colon cancer HCT116 cells were divided into 3 groups. N, N-dimethylsphingosine(DMS) was used to suppress the activity of SphK1. PF573228 was used to suppress the activation of FAK. The cells treated with equal volume of culture medium severed as control group. The cell viability was measured by MTT assay. The protein expression of SphK1, FAK and the EMT relative protein E-cadherin, N-cadherin, vimentin and matrix metalloproteinase(MMP) 2 was analyzed by Western blot. The mRNA expression of SphK1, sphingosine-1-phosphate(S1P), FAK, E-cadherin and vimentin was detected by real-time PCR. The ability of tumor cell migration was measured by wound-healing assay. RESULTS: The cell viability of HCT116 cells was suppressed by DMS and PF573228 in dose and time dependent manners. DMS significantly suppressed the expression of SphK1, FAK, N-cadherin, vimentin and MMP2, meanwhile enhanced the expression of E-cadherin. PF573228 reduced the expression of FAK, SphK1, N-cadherin, vimentin and MMP2, meanwhile increased the expression of E-cadherin(P<0.01). In addition, the migration ability of HCT116 cells was significantly decreased by treating with DMS and PF573228(P<0.01). Compared with control group, the mRNA expression of FAK, SphK1, S1P and vimentin was decreased, while the expression of E-cadherin was increased significantly in PF573228 group and DMS group(P<0.05). CONCLUSION: SphK1 and FAK signaling pathways may play an important role in the occurrence of EMT in the colon cancer HCT116 cells.     

Effect of curcumin on ox-LDL-induced HAEC injury

AIM: To investigate the effect of curcumin on oxidized low-density lipoprotein (ox-LDL)-induced injury of human aortic endothelial cells (HAECs). METHODS: HAECs were pre-treated with curcumin at different concentrations and then treated with ox-LDL. The cell viability was assessed by MTT assay. The cell proliferation ability was analyzed by EdU assay. ELISA was used to determine the concentrations of interleukin-6 (IL-6), transforming growth factor β1 (TGFβ1), high mobility group box-1 protein (HMGB1) and secretory receptor for advanced glycation end products (sRAGE) in the HAEC culture medium. The binding activity of peroxisome proliferator-activated receptor γ (PPARγ) was evaluated by electrophoretic mobility shift assay. The protein levels of HO-1, HMGB1, RAGE,IL-6,TGFβ1 and phosphorylated PPARγ in the HAECs were determined by Western blot. RESULTS: The viability and the proliferation ability decreased significantly in the HAECs treated with ox-LDL. The PPARγ/HO-1 signaling pathway was inhibited while its down-stream HMGB1/RAGE signaling pathway was activated by ox-LDL. The levels of IL-6, TGFβ1, HMGB1 and sRAGE were increased. Pre-treatment with curcumin activated PPARγ/HO-1 signaling pathway and inhibited HMGB1/RAGE signaling pathway in ox-LDL treated HAECs in a concentration-dependent manner. The levels of IL-6, TGFβ1, HMGB1 and sRAGE were also decreased dramatically by pre-treatment of curcumin in a concentration-dependent manner. CONCLUSION: ox-LDL induces HAEC damage by inhibiting PPARγ/HO-1 to activate HMGB1/RAGE inflammatory signaling. Curcumin exerts protective effect on ox-LDL treated HAECs via activating PPARγ/HO-1 signaling pathway.     

Activation of smad signaling and collagenⅠsynthesis in NRK52E cells induced by advanced glycation end products

AIM:To investigate the effects of advanced glycation end products on activation of Smad signaling pathway and collagenⅠ synthesis in proximal tubular epithelial cells. METHODS:Advanced glycation end products (AGE-BSA) were prepared by incubation of bovine serum albumin (BSA) with D-glucose. Normal rat proximal tubular epithelial (NRK52E) cells were cultured in RPMI-1640 medium with AGE-BSA. Phosphorylation and nuclear translocation of Smad2/3 were examined by immunocytochemistry. Levels of TGF-β1 in supernatant of cell culture were measured by enzyme-linked immunosorbent assay (ELISA). Expression of TGF-β1, Smad2, Smad3 and Smad7 mRNA were detected by RT-PCR. Expression of α-SMA , E-cadherin and collagenⅠproteins were detected by Western blotting.RESULTS:AGE-BSA induced Smad2/3 phosphorylation and nuclear translocation, two peaks occured at 30 min (68% vs 16%, P<0.05) and 24 h (76% vs 16%, P<0.05) compared to 0 min. The level of TGF-β1 markedly increased in supernatant of cell culture by induced AGE-BSA at 24 h and 48 h. The expression of TGF-β1 mRNA markedly increased at 24 h, and associated with high expression of Smad2, Smad3 and Smad7 mRNA at 48 h. AGE-BSA up-regulated significantly the expression of α-SMA and collagenⅠproteins, down-regulated the expression of E-cadherin protein. CONCLUSION:AGEs induces activation of Smad signaling, as well as transdifferentiation and collagenⅠ synthesis in proximal tubular epithelial cells.     

Promoting cell viability and migration of gastric cancer cells by PRRX2 via activation of Wnt/β-catenin signaling pathway

AIM: To study the effect of paired-related homeobox 2 (PRRX2) gene on the viability and migration ability of gastric cancer cells, and to analyze the underlying mechanism of regulating Wnt/β-catenin signaling pathway.METHODS: The expression of PRRX2 in gastric cancer and normal gastric tissue and the correlation between PRRX2 expression in gastric cancer tissues with the overall survival rate of gastric cancer patients were analyzed by bioinformatics. The small interfering RNA (siRNA) and over-expressed plasmids of PRRX2 were transfected into gastric cancer cells MGC-803 and SGC-7901, respectively. MTT assay and Transwell assay were used to detect the viability and migration ability of gastric cancer cells. Western blot and TOPflash/FOPflash dual-luciferase reporter gene assay were used to detect the activity of Wnt/β-catenin signaling pathway. Co-immunoprecipitation was used to detected the interaction between PRRX2 and β-catenin proteins.RESULTS: Knockdown of PRRX2 attenuated the viability and migration ability of gastric cancer cell line MGC-803 (P<0.05). Over-expression of PRRX2 enhanced the viability and migration ability of SGC-7901 cells (P<0.05), increased the protein levels of β-catenin, c-Myc and cyclin D1 (P<0.05) and the activity of TOPflash/FOPflash dual-luciferase reporter gene (P<0.05). PRRX2 interacted with β-catenin protein in gastric cancer cells.CONCLUSION: PRRX2 promotes the viability and migration ability of gastric cancer cells, which may be related to Wnt/β-catenin signaling pathway.     

Effects of propofol on invasion and migration of human gastric cancer SGC-7901 cells

AIM: To investigate the effects of propofol on invasion and migration of gastric cancer cell line SGC-7901. METHODS: Cultured gastric cancer cell line SGC-7901 was randomly divided into 4 groups, and then diffe-rent concentrations (1, 3, 5 and 7 mg/L) of propofol were added and incubated for 24 h. The cell viability was measured by MTT assay. The invasion and migration abilities of the SGC-7901 cells were detected by Transwell assay and wound-healing assay. The expression of cysteine-rich angiogenic inducer 61 (CYR61), CD44v6 and matrix metalloproteinase-7 (MMP-7) in the SGC-7901 cells were examined by immunocytochemistry and Western blot.RESULTS: Propofol at 5 mg/L does not affect the viability of SGC-7901 cells, whereas significantly suppresses the invasion and migration abilities, and down-regulates the expression of CD44v6 and MMP-7 (P<0.05). CONCLUSION: The decreased invasion and migration abilities of SGC-7901 cells were partly due to the inhibition of CD44v6 and MMP-7 expression.     

miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.     

Effects of NOB1 gene silencing on drug sensitivity and invasion and migration abilities of human colon cancer SW480 cells

AIM:To investigate the effect of NOB1 gene expression knock-down by transfection of small interfering RNA (siRNA) on the viability, drug sensitivity, apoptosis, cell cycle distribution, and invasion and migration abilities of human colon cancer SW480 cells. METHODS:NOB1 siRNA was transfected into SW480 cells using Lipofectamine 3000. The mRNA and protein levels of NOB1 in the SW480 cells were determined by real-time PCR and Western blot. The cell viability and sensitivity to different chemotherapeutic drugs (cisplatin, 5-fluorouracil, oxaliplatin and capecitabine) were detected by MTT assay after knock-down of NOB1 gene expression in the SW480 cells. The apoptosis and cell cycle distribution of SW480 cells were analyzed by flow cytometry. The invasion and migration abilities of SW480 cells were detected by Transwell assay. RESULTS:After transfection with NOB1 siRNA, the mRNA and protein levels of NOB1 in the SW480 cells were significantly decreased (P<0.05). Compared with control group and control siRNA group, the viability of SW480 cells in NOB1 siRNA group was significantly decreased at 24~72 h. The half maximal inhibitory concentrations of the chemotherapy drugs cisplatin, 5-fluorouracil, oxaliplatin and capecitabine were significantly decreased. The apoptotic rate was significantly increased and the cell cycle were blocked. The cell invasion and migration abilities were significantly reduced (P<0.05). CONCLUSION:Knock-down of NOB1 gene expression inhibits the viability and invasion and migration abilities of colon cancer SW480 cells, and promotes drug sensitivity and apoptosis. NOB1 may be a new target for diagnosis and treatment of colon cancer.