RNA干扰技术对SKOV3细胞EGFR表达的抑制作用

目的探讨RNA干扰(RNAi)技术对卵巢腺癌细胞株SKOV3表皮生长因子受体(EGFR)表达的抑制作用。方法体外化学合成EGFR序列特异性双链RNA(dsRNA),用lipofectamin 2000转染SKOV3细胞,采用RT-PCR和Western blotting技术检测EGFR mRNA和蛋白水平的变化,通过水溶性四唑盐(WST-1)比色法检测细胞增殖力,采用细胞黏附和移动实验检测细胞体外黏附和移动能力,通过细胞侵袭重建基底膜实验检测细胞体外侵袭能力。结果序列特异性dsRNA-EGFR对SKOV3细胞EGFR mRNA和蛋白的抑制率分别为73.65%和58.94%。dsRNA-EGFR组在转染后24、48、72 h细胞增殖抑制率分别为22.54%、35.76%、31.07%;dsRNA-EGFR组在30 min和90 min黏附实验中的黏附率分别下降12.11%和18.66%;转染dsRNA-EGFR后细胞体外移动和侵袭能力的抑制率分别为25.47%和22.08%。结论化学合成的dsRNA可抑制卵巢癌SKOV3细胞EGFR表达,抑制细胞增殖、移动、黏附和侵袭力。

Suppression effect of RNA interference on expression of EGFR of SKOV3 cell line

Objective To investigate the suppression effect of RNA interference(RNAi) on the expression of epidermal growth factor receptor(EGFR) of ovarian cancer SKOV3 cell line. Methods SKOV3 cells were transfected with synthetic EGFR sequence-specific dsRNA by lipofectamin 2000.The expression level of EGFR mRNA and protein were detected by RT-PCR and Western blotting,respectively.The proliferation of transfected SKOV3 cells was detected by WST-1 cell proliferation assay.The abilities of adhesion,migration and invasion were detected by cell adhesion assay,cell migration assay and Matrigel invasion assay,respectively. Results The suppression rates on EGFR mRNA and protein by sequence-specific dsRNA-EGFR were 73.65% and 58.94%,respectively.The cell proliferation was inhibited by 22.54%,35.76% and 31.07%,respectively at 24,48 and 72 h after transfection.The adhesion of sequence-specific dsRNA transfected SKOV3 cells for 30 min and 90 min were reduced by 12.11% and 18.66%,respectively.The migration and invasion of SKOV3 cells were decreased by 25.47% and 22.08%,respectively. Conclusion The down-regulation of EGFR expression of ovarian cancer by sequence-specific dsRNA can lead to the suppression of proliferation,adhesion,migration and invasion of ovarian cancer cells.