Abstract:
Objective To investigate the effect of silencing MFG-E8 gene on the sensitivity of SKOV3 cells to anticancer drugs and related mechanisms.
Methods SKOV3 cells were transfected with MFG-E8 siRNA (Msi) and NC siRNA (Csi), respectively and the efficiency of transfection was confirmed by Western blot. The sensitivity of SKOV3 cells to cisplatin was observed by CCK-8 assay after transfection. The mRNA expression of ABCB1 and ABCC1 were detected by qRT-PCR. Effect of silencing MFG-E8 on the expression of ETM-related protein was detected by qRT-PCR and Western blot.
Results MFG-E8 siRNA could effectively silence the expression of MFG-E8 protein. With the increasing drug concentration, the proliferation inhibition rate of each group also increased, and the cell proliferation inhibition rate of MFG-E8 siRNA group increased significantly (P < 0.01). Compared with NC siRNA group, downregulation of MFG-E8 expression led to decreased SKOV3 cell proliferation at 48h or 72h after 3 μg/ml cisplatin treat ment (P < 0.05). qRT-PCR results showed that the mRNA expression of ABCB1 and ABCC1 in Msi group were significantly lower than those in Csi group. qRT-PCR and Western blot results showed that silencing MFG-E8 gene down-regulated the mRNA and protein expression of N-Cadherin, Vimentin and Snail and up-regulated the expression of E-Cadherin.
Conclusion Silencing the MFG-E8 gene can increase the sensitivity of SKOV3 cells against anti-tumor drugs and down-regulate the mRNA expression of ABCB1 and ABCC1, which may be related to the inhibition of EMT progression.
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