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目的:探讨RNA干扰技术沉默SRC-1基因后对PC-3细胞生长及侵袭性的影响。方法:将设计好的siRNA,用脂质体法转染PC-3细胞,通过Q-PCR、蛋白免疫印迹检测SRC-1的表达变化,并用CCK-8法检测细胞生长情况,用transwell小室检测PC-3细胞侵袭力。结果:转染SRC-1siRNA24小时后的前列腺癌PC-3细胞系中SRC-1mRNA的相对表达量下降约42%,48小时后下降约79%,差异具有统计学意义(P<0.05);转染24小时、48小时后PC-3细胞SRC-1蛋白的表达量(24h 0.5536±0.071、48h 0.3419±0.025)与阴性对照组(24h 0.8562±0.092、48h 0.8791±0.076)、转染试剂组(24h 0.7992±0.072、48h 0.9731±0.051)和空白对照组(24h 0.8375±0.054、48h 0.8826±0.043)相比明显减少(P<0.05)。在转染24小时、48小时、72小时、96小时后PC-3细胞生长情况(吸光度A值:24h 0.324±0.0252、48h 0.689±0.0141、72h 1.0032±0.0166、96h 1.4650±0.0327)与阴性对照组(24h 0.3216±0.0152、48h 0.7014±0.017、72h 0.9902±0.0272、96h 1.4596±0.0141)、转染试剂组(24h 0.3222±0.0343、48h 0.6904±0.0301、72h 0.993±0.0383、96h 1.4574±0.0464)和空白对照组(24h 0.3316±0.0192、48h0.7092±0.0265、72h 1.0136±0.0130、96h 1.4596±0.0128)比较没有明显变化(P>0.05),但是PC-3细胞的侵袭能力明显下降,干扰实验组的细胞(38±9.01)与阴性对照组(83.33±10.78)、转染试剂组(83.67±10.016)及空白对照组(84.67±11.24)相比较,穿过小室膜的细胞数量明显减少(P<0.05)。结论:siRNA有效地阻断了PC-3细胞中SRC-1基因的表达,SRC-1基因在mRNA和蛋白水平上的表达明显下调(P<0.01),并使转染后的PC-3细胞的侵袭能力下降,但转染后PC-3细胞生长无明显变化。
Objective: To investigate the effect of RNA interference on the growth and invasiveness of PC-3 cells after SRC-1 gene silencing. Methods: The designed siRNA was transfected into PC-3 cells by lipofectamine. The expression of SRC-1 was detected by Q-PCR and Western blotting. The cell growth was detected by CCK-8 assay, PC-3 cell invasiveness. Results: The relative expression level of SRC-1 mRNA in prostate cancer PC-3 cells transfected with SRC-1 siRNA decreased by about 42% and decreased by about 79% after 48 hours. The difference was statistically significant (P <0.05) The expression of SRC-1 protein in PC-3 cells after 24h and 48h treatment was significantly higher than that in control group (24h 0.8562 ± 0.092,48h 0.8791 ± 0.076) 24h, 0.7992 ± 0.072, 48h, 0.9731 ± 0.051) and control group (24h 0.8375 ± 0.054,48h 0.8826 ± 0.043). The growth of PC-3 cells at 24 hours, 48 hours, 72 hours and 96 hours after transfection (absorbance A value: 24h 0.324 ± 0.0252,48h 0.689 ± 0.0141,72h 1.0032 ± 0.0166,96h 1.4650 ± 0.0327) were compared with the negative control group (24h 0.3216 ± 0.0152,48h 0.7014 ± 0.017,72h 0.9902 ± 0.0272,96h 1.4596 ± 0.0141), transfection reagent group (24h 0.3222 ± 0.0343,48h 0.6904 ± 0.0301,72h 0.993 ± 0.0383,96h 1.4574 ± 0.0464) and blank control Group (24h 0.3316 ± 0.0192,48h0.7092 ± 0.0265,72h 1.0136 ± 0.0130,96h 1.4596 ± 0.0128) showed no significant change (P> 0.05), but the invasive ability of PC-3 cells was significantly decreased, 38 ± 9.01). Compared with the control group (83.33 ± 10.78), transfection reagent group (83.67 ± 10.016) and blank control group (84.67 ± 11.24), the number of cells passing through the ventricular membrane decreased significantly (P <0.05). CONCLUSIONS: siRNA effectively blocked the expression of SRC-1 gene in PC-3 cells. The expression of SRC-1 gene at mRNA and protein levels was significantly down-regulated (P <0.01), and the transfection of PC-3 cells The invasive ability of PC-3 cells did not change significantly after transfection.