| Background and Objective：Ovarian cancer is a common one of malignant tumors in female feproductivesystem. Ovarian cancer has the most admonishing total prognosis due to diagnosis atlate stage, its insidious onsets, concealed primary focus, relapse, tendency to developchemotherapy resistance and dissemination. it has the highest death rate and is boundto generate severe consequences for the health and lives of women in allgynecological tumors. the five year survival rate of ovarian cancer is30%. In recentyears, despite improvements in the molecular mechanism of ovarian cancer, whichgained thumping developments in mechanism of chemotherapy resistance, theimportant molecular mechanism of ovarian cancer is still indistinct.Metastasis-associated in colon cancer-1(MACC1) was found out bygenomewide expression analytical technique in metastasis focus and primary focus ofcolon cancer. The expressions of MACC1were significantly higher in gastric cancer,lung cancer, liver cancer and pancreatic cancer, MACC1could promote the growth,adhesion, invasion and migration of many tumor cells. Hepatocyte growth factor(HGF) could promote the appreciation, conglutination and invasion of tumour cells,and play an important role in the appreciation, metastasis and invasion of many malignant tumour cells, including ovarian cancer, according to statistics, MACC1wasproved as a significant regulator of MET in HGF/C-met signaling. At present, howabout MACC1overexpression in SKOV-3/DDP, how about the mechanism ofMACC1overexpression induced SKOV-3/DDP cells to enhance sensitive of DDP.To examine the enhancer of metastasis-associated in colon cancer-1（MACC1）gene expression in cisplatin-resistant ovarian cancer cell line SKOV-3/DDP and itsparental cell line SKOV-3and to investigate whether silencing MACC1by RNAinterference could reverse cisplatin resistance of SKOV-3/DDP cells.Materials and Experimental Methods:1. Expressions of MACC1mRNA and protein in cells were detected byreal-time in quantitative and Westem blotting respectively.2. RT-PCR and Westem blotting were used to validate whether the MACC1gene expression was silenced.3. The effect of cisplatin on cell growth was evaluated using MTT assays.4. The influence of silencing MACC1are explored on chemoresistance by flowcytometry analysis.5. Statistical analysis: The SPSS statistical package program17.0was appliedto analyze the results of experiment. The level of expression among different tissueswere tested by One-way analysis of variance. The LSD test was used in two groups. P<0.05was considered statistically significant.Results:1. The MACC1mRNA and protein expression levels in SKOV-3/DDP cellswere (2.66±0.54) and (1.95±0.45) folds of those in SKOV-3cells. The difference wassignificant (P<0.05).2. he expression levels of metastasis-associated in colon cancer-1(MACC1)mRNA and protein in ovarian cancer cell line SKOV3/DDP after transfection withsmall hairpin RNA targeting MACC1gene (MACC1-shRNA) were detected byreverse transcription-PCR (RT-PCR) and Western blotting, respectively. SKOV3/ DDP: Without transfection as the control group; SKOV-3/DDP-shVector:Transfected with empty plasmids as the negative control group; SKOV-3/DDP-shMACC1: Transfected with the combinant plasmids contained with MACC1-shRNA as the experimental group. The expression levels of MACC1mRNA andprotein in SKOV-3/DDP-shMACC1cells were lower than those in SKOV-3/DDP-shVector cells and SKOV-3/DDP cells (P<0.05, n=3).3. The proliferation viabity of ovarian cancer SKOV3/DDP cells transfectedwith MACC1-shRNA and treated with cisplatin (DDP, at10,20,30,40,50and60μmol/L) for48h were detected by MTT method (n=3). SKOV3/DDP: Withouttransfection as the blank control group; SKOV-3/DDP-shVector: Transfected withempty plasmids as the negative control group; SKOV-3/DDP-shMACC1: Transfectedwith the combinant plasmids contained with MACC1-shRNA as the experimentalgroup. The sensitivity of SKOV3/DDP cells transfected with MACC1-shRNA toDDP was enhanced as compared with that of SKOV3/DDP cells untransfected andtransfected with empty vectors.4. The effect of MACC1gene expression down-regulation on apoptosis rate ofovarian cancer SKOV3/DDP cells was checked by flow cytometry. SKOV3/DDP:Without transfection as the blank control group; SKOV-3/DDP-shVector: Transfectedwith empty plasmids as the negative control group; SKOV-3/DDP-shMACC1:Transfected with the combinant plasmids contained with MACC1-shRNA as theexperimental group. The apopototic rate of SKOV-3/DDP-shMACC1cells wassignificantly higher than that in SKOV-3/DDP-shVector and SKOV-3/DDP cells(P<0.05, n=3).Conclusions:1. MACC1is overexpressed in SKOV-3/DDP cell line compared with parentalcell line SKOV-3.2. Silencing MACC1by RNA interference effectively reverse the resistance ofSKOV-3/DDP cells to cisplatin. |
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