The Effect Of SiRNA Silencing MACC1 On The Proliferation And Invasion Of Hela Cells

In recent years,cervical cancer has become the most common gynecologic malignant tumor in the clinic,which poses a serious threat to the life and health of the women.There is an urgent need to understand the pathogenesis of cervical cancer and the means of gene regulation related factors.HeLa cells are one of the cervix cancer cells,which are often used to study the pathogenesis of cervical cancer.Colon cancer metastasis associated gene 1(metastasis-associated in colon cancer-1,MACC1)was originally that is just one of the important genes associated with colon cancer,after study in recent years,the study found that MACC1 can affect many tumors including ovarian cancer cells,lung cancer,liver cancer and other proliferative ability or to the surrounding tissue invasion,thereby affecting the development and metastasis of tumor.However,the expression level of MACC1 in cervical cancer HeLa cell proliferation and invasiveness and silencing of MACC1 cervical cancer HeLa cells is unclear,the research will be conducted on the proliferation of MACC1 and cervical cancer and the correlation of the invasiveness of discussion,to fill the research gap in this area.Purpose:To explore the expression level of MACC1 in HeLa cells and the effect of silencing of MACC1 on the proliferation and invasiveness of HeLa cells.Methods:(1)the expression of MACC1 RT-PCR detection of HeLa cells and normal cells;(2)select siRNA silencing sequence;(3)HeLa cells were cultured in normal cells,HeLa cells and MACC1 silencing;(4)siRNA after transfection,using Western blotting method to detect the expression of MACC1 protein in HeLa cells;(5)si RNA after transfection,using MTT method to detect the change of proliferation ability of HeLa cells;(6)Western blotting detection of siRNA E-cadherin after transfection and the expression level of N-cadherin;(7)Transwell migration assay in vitro.Results:(1)The relative expression level of MACC1 in normal cells was 1.612,and therelative expression level of MACC1 in HeLa cells was 4.625,compared with the obvious statistical difference(P < 0.001).(2)the siRNA sequence 5’-GUGCUCAGGACUGAUGAAATT-3 can effectively inhibit MACC1 expression;(3)after the expression of MACC1 gene was transfected with si RNA,the cell proliferation rate was only 71 % of normal HeLa cells and control cells.(4)the expression level of e-cadherin was up-regulated and the expression level of n-cadherin was down-regulated after adding siRNA in the culture medium;(5)after transfection of siRNA,the number of cells in the Tranwell chamber was significantly lower than that of HeLa cells without transfection of siRNA.Conclusions:(1)the expression level of MACC1 in HeLa cells was significantly higher than that of normal cells;(2)si RNA silencing sequence 5 ’-GUGCUCAGGACUGAU GAAATT-3’(expression interference sequence)inhibits the expression of MACC1,which can be used in MACC1 silencing related experiments;(3)after the expression of MACC1 was silenced,the proliferation rate of HeLa cells decreased.(4)the expression of MACC1 plays an enhanced role in the invasion capacity of cervical cancer tissue cells;(5)MACC1 silence inhibits the migration of cervical cancer(HeLa cells),and the presence of MACC1 in HeLa cells can promote the migration of HeLa cells.