Effect of ginsenoside Rh2 on cell proliferation and cytoskeleton in hepatocarcinoma SMMC-7721 cells

AIM: To investigate the changes of cell growth and cytoskeleton in hepatocarcinoma SMMC-7721 cells treated with ginsenoside Rh₂.METHODS: Cell viability and apoptosis under the conditions of ginsenoside Rh₂ exposure at different concentrations were measured by MTT test and flow cytometry,respectively. The morphological changes of F-actin labeled with FITC-phalloidin were observed under confocal laser scanning microscope. The structures of nuclear matrix-intermediate fibre system were observed under transmission electron microscope (TEM).RESULTS: Rh₂ at 40 mg/L for 4 days inhibited the proliferation and induced apoptosis in SMMC-7721 cells more than those in control group, 10 mg/L Rh₂ group and 20 mg/L Rh₂ group. The F-actin in the cells treated with Rh₂ was well-distributed, lined up in order and the number of fibers increased, while those in the control cells were in disorder and punctiform. The results of whole mount TEM indicated that the intermediate fiber was plentiful, well-distributed and interweaved into a regular network in Rh₂ treated cells.CONCLUSION: Rh₂ effectively inhibits the cell proliferation, increases the cell apoptosis and induces the change of the cytoskeleton alignment in SMMC-7721 cells.     

Effects of changing PKC activity on proliferation and telomerase expression in human hepatocellular carcinoma cells

AIM: To investigate whether protein kinase C (PKC) is involved in the proliferation and the telomerase expression in human hepatocellular carcinoma cells. METHODS: Human hepatocellular carcinoma cells (BEL-7402) were treated with exogenous phorbol-12-myristate-13-acetate (PMA, PKC activator) and staurosporine (SP, PKC inhibitor) for 48 hours. The techniques of cell culture and the telomeric repeat amplification protocol silver staining in combination with computer image scanning system in vitro were used to observe the variations of the growth and the telomerase expression. RESULTS: The proliferative potential of BEL-7402 cells was decreased by the action of PMA as well as SP, and the telomerase expression was also inhibited by PMA and SP. CONCLUSION: Our findings suggest that the proliferation of human hepatocellular carcinoma cells and the telomerase expression may be related to PKC.     

Up-regulation of microRNA-187* inhibits proliferation of human colon cancer cell line HCT116

AIM:To investigate the expression of microRNA-187* (miR-187*) in human colon cancer cell lines and normal colon tissues, and to determine the effects of miR-187* up-regulation on the proliferation and cell cycle of human colon cancer cell line HCT116. METHODS:The expression profiling of miRNAs in 3 colorectal adenocarcinoma samples and their matched normal tissue samples was performed using miRNA microarray chip. Total RNA was isolated from 8 colon cancer cell lines and 10 normal colon tissues. The miR-187* level was detected by Taqman real-time RT-PCR. B-cell-specific Moloney murine leukemia virus integration site 1 (BMI-1), the possible target of miR-187*, was also detected. Synthetic miR-187* mimics were transfected into HCT116 cell line by LipofectamineTM 2000. The mRNA expression of miR-187* and BMI-1 in HCT116 cell line was measured by real-time RT-PCR. Cell growth and cell cycle were assayed by MTS method and flow cytometry. RESULTS:miR-187* was found to be differentially expressed between colorectal adenocarcinoma and normal tissues. The expression of miR-187* in 8 colon cancer cell lines was down-regulated, while BMI-1 mRNA was up-regulated. Compared with blank control group, miR-187* expression was remarkably increased after transfection with miR-187* mimics, and ectopic expression of miR-187* significantly inhibited the mRNA expression of BMI-1. The cell growth was inhibited in miR-187* mimics group, and proliferating cell nuclear antigen (PCNA) mRNA expression was decreased. The cells at G2/M phase in miR-187* mimics group were significantly increased. CONCLUSION: miR-187* is down-regulated in human colon cancer cell lines. Up-regulation of miR-187* not only inhibits the proliferation but also influence the cell cycle of HCT116 cells, which might act as a tumor suppressor in colorectal cancer by inhibiting the expression of BMI-1.     

Effect of SC58125 on cell proliferation and apoptosis in HepG-2 cells

AIM: To clarify the effect of SC58125 on cell proliferation and apoptosis in HepG-2 cells and explore the molecular mechanisms. METHODS: Cell culture, MTT, TUNEL, DNA ladder, flow cytometry and Western blot analysis were employed in the present study. RESULTS: SC58125 inhibited the growth of HepG-2 cells and induced the apoptosis. Furthermore, it arrested G₀/G₁ phase and inhibited S phase in HepG-2 cells. Depressed expression of P33^cdk2, P34^cdc2, cyclin B₁, cyclin E, Mpm-2, Rb, PCNA proteins were found in HepG-2 cells treated with SC58125. CONCLUSION: SC58125 inhibits cell proliferation and induces apoptosis, which may be related to the altered low protein levels of P33^cdk2, P34^cdc2, cyclin B₁, cyclin E, Mpm-2, Rb, PCNA.     

Effect of antisense RNA targeting Polo-like kinase 1 on cell growth in A549 lung cancer cells

AIM: To investigate the effect of Polo-like kinase-1 (Plk1) depletion on cell cycle progression and cell growth in lung cancer cells.METHODS: A recombinant plasmid containing antisense RNA targeting Plk1 (pcDNA3-Plk1) was transfected into A549 cells by lipofectine. RT-PCR and Western blotting were used to examine Plk1 gene expression. Cell proliferation was evaluated by cell counting and BrdU labeling. Cell cycle distribution and apoptosis were examined by flow cytometry. Inhibition rate (IR) of vinorebline (NVB) was determined by MTT assay. RESULTS: After transfected with pcDNA3-Plk1 into A549 cells, the expression levels of Plk1 mRNA and protein were greatly decreased. Abnormal morphological changes of cells and growth inhibition were observed in pcDNA3-Plk1 transfected cells. The BrdU labeling index was significantly lower than that in control group (P<0.05). Cells showed a strong G₂/M arrest and apoptosis 72 h post transfection. IR of vinorebline in pcDNA3-Plk1 transfected groups was significantly higher than that in other groups. CONCLUSION: Antisense RNA targeting Plk1 is capable of suppressing Plk1 expression, and therefore, significantly inhibits cellular proliferation, induces cell cycle arrest and apoptosis. Moreover, the sensitivity of lung cancer cells to chemotherapy is increased.     

Expression of microRNA-1284 in gastric cancer and underlying mechanism

AIM: To evaluate the correlation between microRNA-1284 (miR-1284) and gastric cancer, and to investigate the underlying mechanism. METHODS: The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer (GC) tissue samples and 63 non-malignant adjacent tissue samples. The correlation between miR-1284 and the clinicopathological feature of GC was analyzed. Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells. After transfection, the expression of miR-1284 was examined by real-time PCR. The cell activity was evaluated by CCK-8 assay. The cell cycle and apoptosis were determined by flow cytometry. The ability of cell migration was measured by wound-healing assay. The potential target gene of miR-1284 was predicted by online bioinformatic softwares. The expression of JAG1 mRNA was examined by real-time PCR. The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting. RESULTS: Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples (P<0.05). The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases (P>0.05), but significantly associated with histologic grading (P<0.05). Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly increased (P<0.05), the percentages of apoptotic cells and the cells in G₀/G₁ phase were significantly increased (P<0.05), the cells activity and ability of migration were significantly decreased (P<0.05), and the expression of JAG1, Notch1 and NF-κB was significantly decreased (P<0.05). CONCLUSION: The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.     

Effect of tanshinone IIA on expression of cell cycle regulators and proliferation of pancreatic cancer cells

AIM：To observe the effect of tanshinone IIA on the expression of cell cycle regulators and the proliferation of pancreatic cancer cell line BX-PC-3. METHODS： The pancreatic cancer cell line BX-PC-3 was treated with tanshinone ⅡA at various concentrations for 48 h. The inhibition of proliferation was measured by MTT method. The change of the cell cycle was detected by flow cytometry. The protein levels of cyclin A and cyclin D2 were determined by Western blotting. RESULTS：Tanshinoone IIA significantly inhibited the proliferation of BX-PC-3 cells in a dose-dependent manner. The cancer cells were arrested in stage G0/G1 after treated with tanshinone IIA at low dose. The protein levels of cyclin A and cyclin D2 were decreased after drug intervention. CONCLUSION：Tanshinone IIA inhibits the proliferation of human pancreatic cancer cell line BX-PC-3 and the expression of cell cycle-promoting factors (cyclin A and cyclin D2), which may be the mechanism of attenuating the proliferation of pancreatic cancer cells.     

Mechanism of microRNA-138-5p inhibiting proliferation,migration and invasion of lung cancer cells

AIM: To investigate the mechanism of microRNA-138-5p (miR-138-5p) inhibiting the proliferation, migration and invasion abilities of lung cancer cells.METHODS: The lung cancer A549 and H460 cells were transfected with miR-NC (control group) or miR-138-5p (experimental group). The bioinformatic analysis was performed to predict the target genes of miR-138-5p.The expression levels of miR-138-5p, forkhead box protein C1 (FOXC1) mRNA and vimentin mRNA were detected by RT-qPCR. The protein expression of FOXC1, vimentin, E-cadherin, N-cadherin and β-catenin was determined by Western blot. MTS method and colony formation assay were used to detect cell viability and proliferation ability. Wound healing assay and Transwell assay were used to detect cell migration and invasion ability.RESULTS: Over-expression of miR-138-5p significantly reduced the expression of FOXC1 and vimentin at mRNA and protein levels (P<0.05). The expression of E-cadherin and β-catenin were up-regulated and the expression of N-cadherin was down-regulated. The proliferation, migration and invasion abilities of the lung cancer cells were inhibited by the over-expression of miR-138-5p.CONCLUSION: miR-138-5p inhibits the proliferation, migration and invasion abilities of lung cancer cells by targeting FOXC1 and vimentin. It may be a potential target for lung cancer gene therapy.     

Effect of protein kinase C-α antisense oligonucleotide on cell cycle of CNE-2Z cells

AIM:To observe the effect of protein kinase C-α(PKCα)antisense oligonucleotide on cell growth, cell cycle and the expression of cyclin E in human poor-differentiated nasopharyngeal carcinoma(NPC) cell line CNE-2Z. METHODS:Antisense PKCα was transfected by cationic liposome(LP) in CNE-2Z cells to analyze the cell growth and cell cycle by MTT colorimetric assay and flow cytometry, respectively. Moreover, the expression of cyclin E was determined by immunocellularchemistry and scanning the result of dot-blotting. RESULTS:①With the concentration of antisense PKCα increasing, the relative cell growth index was decreased gradually(P＜0.01). ②After treated with antisense PKCα, the percentage of cells in G₁ phase enhanced(P＜0.05). ③Compared with the control group, the expressing intensity of cyclin E reduced in antisense PKCα group, and the expression of cyclin E decreased to 66.5%±18.4％(P＜0.05) of the control by scanning quantitative analysis. CONCLUSION:These results indicated that antisense PKCα may inhibit cell growth in CNE-2Z via suppressing the expression of cyclin E and hindering cell process in G₁ phase.     

and laminin on adhesion and proliferation of human hepatocellular carcinoma cell

AIM:To explore the effects of PMA(phorbol-12-myristate-13-acetate, a tumor promoter, mimicking the action of diacylglycerol on PKC)and laminin on the adhesion and the proliferation of human hepatocellular carcinoma cells, and provide a new clue to liver cancer treatment.METHODS:Human hepatocellular carcinoma cell line(BEL-7402)was used to identify the endogenous laminin and protein kinase C-α(PKC-α) expression, and the effects of laminin and PMA on the adhesion and the proliferation were also investigatedin vitro.RESULTS:By the effect of exogenous laminin, human hepatocellular carcinoma cell (BEL-7402) possessed endogenous laminin expression and increased the adhesion and the proliferation, which was showed the synergistic action by the effect of PMA in combination. By the action of PMA alone, the proliferation and the PKC-α expression increased by exogenous laminin were decreased, and the adhesion and the endogenous laminin expression were increased.CONCLUSIONS:The finding suggested that the adhesion and the proliferation of human hepatocellular carcinoma cell were closely related to the effects of endogenous or exogenous laminin, which were associated with cPKC-α activity. Therefore, the application of anti-laminin antibody in combination with PKC antagonist might be a new clue to find out the therapy for liver cancer.     

Effect of lentiviral vector of microRNA-9-1 on differentiation of mouse bone marrow mesenchymal stem cells into neurons

AIM: To investigate the role of microRNA-9 in inducing bone marrow mesenchymal stem cell(MSCs) differentiation into neurons.METHODS: The lentiviral vector of microRNA-9-1(microRNA-9-1-LV) was constructed and transfected into mouse MSCs. The cells were divided into non-transfected group, transfected group(transfected with microRNA-9-1-LV) and negative control group(transfected with FU-RNAi-NC-LV). MSCs were treated with β-mercaptoethanol(β-ME) as an inducer for triggering the cells to differentiate into neurons. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope. The mRNA expression of microtublin-associated protein 2(MAP-2) was detected by RT-PCR. The expression of neuron-specific markers,neuron-specific enolase(NSE), MAP-2 and glial fibrillary acidic protein(GFAP), were measured by immunocytochemical method. The viability of MSCs was determined by MTT method. RESULTS: The results of PCR confirmed successful construction of mouse microRNA-9-1-LV. The virus titer was 1×10¹² TU/L(TU, transduction unit). The best transfection efficiency(up to 91.3%±4.2%) and survival rate appeared when multiply of infection(MOI)was 20 and on 4th day. β-ME induced MSCs to differentiate into neurons and the best efficiency of the induction was observed in transfected group. The expression levels of NSE and MAP-2 in transfected cells were higher than those in the cells of other group(P<0.05).CONCLUSION: MicroRNA-9-1-LV has high transfection efficiency in mouse MSCs. Higher differentiation rate from mouse MSCs to neurons is induced by β-ME after the cells are transfected with microRNA-9-1-LV.     

Effect of miR-375 on viability,cell cycle and apoptosis of colon cancer HCT116 cells

AIM: To investigate the effect of microRNA-375 (miR-375) on the viability, cell cycle and apoptosis of HCT116 cells.METHODS: The expression of miR-375 in different colorectal cancer cell lines was detected by real-time PCR. The miR-375 mimics was transfected into HCT116 cells by Lipofectamine^TM 2000. The mRNA expression of miR-375 and AEG-1 was detected by real-time PCR. The HCT116 cell viability was detected by MTT assay. The changes of apoptosis and cell cycle distribution were analyzed by flow cytometry.RESULTS: Real-time PCR showed that miR-375 expression was the lowest in HCT116 among 4 colorectal cancer cell lines. The expression level of miR-375 significantly increased in miR-375 mimics group compared with that in the negative control group. The high expression level of miR-375 significantly inhibited the mRNA expression of AEG-1. After transfection with miR-375 mimics, the cell viability was inhibited, the apoptotic rate was increased, the proportion of G₁-stage cells was increased, and the proportion of S-stage cells was decreased.CONCLUSION: miR-375 inhibits the viability, mediates the cell cycle arrest and promotes the apoptosis of colon cancer HCT116 cells. miR-375 may act as a tumor suppressor in colorectal cancer by inhibiting AEG-1.     

Effects of miRNA-25 on proliferation of human esophageal squamous-cell carcinoma cell line TE1

AIM: To investigate the effects and mechanisms of microRNA-25(miRNA-25) on the proliferation of human esophageal squamous-cell carcinoma cell line TE1. METHODS: The abundance of miRNA-25 in different tissues was measured by RT-PCR. After silencing or over-expression of miRNA-25 with mimics or inhibitor in TE1 cells, the cell proliferation, cell cycle distribution and the expression of cyclin E1 and cyclin-dependent kinase 2(CDK2) at mRNA and protein levels were measured by CCK-8 assay, BrdU detection, flow cytometry, RT-PCR and Western blot, respectively. RESULTS: miRNA-25 was prominent in esophageal mucosal tissue and highly expressed in TE1 cells (P<0.05). Over-expression of miRNA-25 increased TE1 cell proliferation, promoted the cell cycle progression and enhanced the entrance of the cells into S phase (P<0.05). Inverse results were obtained after down-regulation of miRNA-25(P<0.05). Furthermore, the expression of cyclin E1 and CDK2 at mRNA and protein levels was significantly increased after over-expression of miRNA-25, but decreased after down-regulation of miRNA-25(P<0.05). CONCLUSION: miRNA-25 enhances cell cycle transition by increasing the expression of cyclin E1 and CDK2, thus accelerating TE1 cell proliferation. This study provides a novel mechanism by which miRNA-25 increases the proliferation of human esophageal squamous-cell carcinoma cell line TE1, suggesting that down-regulation of miRNA-25 may be a potential new therapeutic strategy for treating esophageal squamous-cell carcinoma.     

Regulatory effects of Rock2 on cell cycle checkpoint Cdc25A

AIM: To investigate the regulatory effects of Rho-associated coiled-coil-containing protein kinase 2(Rock2) on the cell cycle checkpoint cell division cycle 25A(Cdc25A). METHODS: The protein expression levels of Rock2 and Cdc25A in 51 pairs of hepatocellular carcinoma and the adjacent tissues were detected by Western blotting. shRock2 plasmids were constructed, selected and stably transfected into hepatocellular carcinoma Huh-7 and HepG2 cells. The protein expression of Cdc25A in the cells was determined by Western blotting. Based on the Rock2 interfering sequences, we designed the primers and changed the 4 indicated bases via site-specific mutagenesis. The Rock2-mutant plasmid was verified by sequencing and was transfected into stable Rock2-knockdown cells. The protein expression of Cdc25A was detected by Western blotting, and the cell proliferation was measured by MTT assay. The protein levels of checkpoint kinase(Chk)1/Chk2 were also detected in stable Rock2-knockdown cells. The interaction between Rock2 and Cdc25A was measured by co-immunoprecipitation, and the co-localization of Rock2 and Cdc25A was detected by confocal laser scanning microscopy. RESULTS: Rock2 and Cdc25A were apparently up-regulated in hepatocellular carcinoma,with a significantly positive correlation. The protein expression of Cdc25A was significantly down-regulated in stable Rock2-knockdown cells. The expression of Chk1 and Chk2 was not changed following knockdown of Rock2. The co-immunoprecipitation results verified that Rock2 bound to Cdc25A. The results of confocal laser scanning microscopy showed that Rock2 and Cdc25A were co-localized in hepatocellular carcinoma cells. CONCLUSION: Rock2 positively regulates the cell cycle checkpoint Cdc25A, which is independent of Chk1/Chk2 and this may provide a new target gene for treatment of hepatocellular carcinoma.     

Effect of abnormal expression of miR-141 on malignant biological behaviors of human hepatocarcinoma cells

AIM: To investigate the expression of microRNA-141 (miR-141) in human hepatocellular carcinoma (HCC) cell line SMMC-7721 and normal hepatocyte line HL-7702, and to analyze the effect of abnormal expression of miR-141 on the malignant biological behaviors of human hepatocarcinoma cells. METHODS: The RNA from SMMC-7721 cells and HL-7702 cells was extracted. SYBR Green real-time PCR was performed to detect the expression of miR-141. Synthetic miR-141 mimic and its negative control were transfected into the SMMC-7721 cells, and miR-141 inhibitor and its negative control were transfected into the HL-7702 cells by the method of Lipofectamine. After transfection, MTS assay and BrdU-ELISA were employed to evaluate the effect of miR-141 on the cell proliferation. Flow cytometry was used to detect cell cycle and apoptosis. The changes of migration ability were investigated by Transwell invasion assay. RESULTS: The expression of miR-141 in the SMMC-7721 cells was significantly lower than that in the HL-7702 cells (P < 0.05). Compared with blank group, Lipofectamine group and negative control group, the proliferation of the SMMC-7721 cells transfected with 25 nmol/L miR-141 mimic was significantly inhibited in a time-dependent manner (P < 0.05). The percentages of G₁ phase cells and early apoptotic rate were significantly increased when miR-141 was up-regulated, but the migration ability was inhibited (P < 0.05). Compared with blank group, Lipofectamine group and negative control group, the proliferation of HL-7702 cells transfected with 50 nmol/L miR-141 inhibitor was significantly increased in a time-dependent manner (P < 0.05). When miR-141 was down-regulated, the percentages of G₁ phase cells and early apoptotic rate were significantly decreased, but the migration ability was enhanced (P < 0.05). CONCLUSION: miR-141 is down-regulated in human hepatocarcinoma cell line. Up-regulation of miR-141 will not only inhibit cell proliferation and migration ability, but also affect the cell cycle and apoptosis of SMMC-7721 cells. miR-141 may function as a tumor suppressor gene during HCC development.     

Effect of ClC-3 siRNA on cell cycle of HeLa cells

AIM: To investigate the roles of ClC-3 chloride channels in the regulation of cell cycle and the relationship between ClC-3 chloride channels and the cell cycle regulators, such as cyclin D1, cyclin-dependent kinase (CDK)4, CDK6, P21 and P27 in the HeLa cells.METHODS: ClC-3 genes were silenced by the siRNA technique in the HeLa cells. The transfection efficiency of ClC-3 siRNA was detected by real-time PCR. The cell cycle distribution was analyzed by the flow cytometry. The protein expression of ClC-3, P21, P27, CDK4, CDK6 and cyclin D1 was determined by Western blot.RESULTS: ClC-3 was knocked down by ClC-3 siRNA in the HeLa cells. Transfection of the cells with ClC-3 siRNA arrested the cells at G₀/G₁ phases, decreased the expression of cyclin D1, CDK4 and CDK6, and increased the expression of P21 and P27.CONCLUSION: ClC-3 plays an important role in the cell cycle of HeLa cells through the G₁-S transition point. ClC-3 may regulate the cell cycle progression by up-regulation of cyclin D1, CDK4 and CDK6 expression and/or by down-regulation of P21 and P27 expression.     

The effect of EBV latent membrane protein 1 on proliferation and cell cycle of nasopharyngeal carcinoma cells

AIM:To investigate the effect of Epstain-Barr virus latent membrane protein 1 (EBV-LMP1) on proliferation and cell cycle of nasopharyngeal carcinoma (NPC) cells. METHODS:The expression of EBV-LMP1 was detected by immunohistochemical method (LSAB). Proliferation of NPC cells was identified by MTT method. Cell cycle percentage was detected by flow cytometry analysis. RESULTS: OD value of EBV-LMP1 expressive NPC cells L-CEN1 was much higher than that of both EBV-LMP1 negative NPC cells V-CEN1 and CNE1 (P＜0.01). Compared with the cell cycle percentage in both V-CNE1 and CNE1, the percentage of G₁ was significantly decreased and the percentage of S was much increased in L-CNE1 (P＜0.01). But no obvious differences were observed in all cell cycle percentage between V-CNE1 and CNE1 (P＞0.05).CONCLUSION:The expression of EBV-LMP1 on NPC cell might cause some change of cell cycle and enhance cell proliferation.     

Effect of RNA interference on Polo-like kinase-1 in A549 cells

AIM: To investigate whether RNA interference(RNAi) induced by small interference RNA(siRNA) could suppress Polo-like kinase-1 (Plk 1) expression and its effects in A549 cells.METHODS: A recombinant plasmid containing siRNA targeting Plk1 (psiRNA-hH1-Plk1) was transfected into A549 cells with Lipofectamine 2000.Expressions of Plk1,cyclin B1 and p53 protein were detected by Western blotting.Cell proliferation was evaluated by direct cell counting,while cell cycle and apoptosis were examined by flow cytometry,and expression of α-tubulin was detected by immunofluorescence.RESULTS: The results demonstrated that sequence specific siRNA targeting Plk1 was capable of suppressing Plk1 expression,and reflecting in lower kinase activity in A549 cells.The level of Plk1 protein was reduced by at least 70% after 48 h of psiRNA-hH1-Plk1 treatment relative to controls.Expressions of cyclin B1 and p53 were increased greatly after Plk1 depletion,and cells showed absence of microtubule polymerization and spindle abnormalities in staining for α-tubulin.Growth inhibition,G₂/M arrest and apoptosis were observed in psiRNA-hH1-Plk1 transfected group.CONCLUSION: All these data suggest that siRNA targeted against human Plk1 may be a valuable tool in cancer therapy.     

Effect of microRNA-132 transfection on lipopolysaccharide-induced inflammation in rat alveolar macrophages

AIM: To investigate the effect of microRNA-132 (miR-132) transfection on the lipopolysaccharide (LPS)-induced inflammation in rat alveolar macrophages. METHODS: The rat alveolar macrophage NR8383 cultured without pyrogen in vitrowere divided into blank control group, negative control group and transfected group. The cells in the 3 groups were transfected with phosphate buffer solution (PBS), Lipofectamine 2000 and synthesized miR-132 mimic respectively. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. Real-time PCR was used to detect the expression of miR-132 in the cells. After NR8383 cells were stimulated with LPS for 6 h, the NF-κB DNA-binding activity was measured by electrophoretic mobility shift assay (EMSA). The expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in NR8383 cells was assayed by Western blotting.RESULTS: After transfection, the expression of miR-132 was significantly higher than that in blank control group and negative control group. The growth of NR8383 cells in transfected group was significantly inhibited compared with blank control group and negative control group (P<0.05). After the cells were stimulated with LPS, the productions of NF-κB, TNF-α and IL-6 in transfected NR8383 cells were decreased compared with blank control group and negative control group (P<0.05).CONCLUSION: Transfection of alveolar macrophages with miR-132 significantly suppresses the cell growth, and inhibits inflammatory responses induced by LPS.