层黏素受体靶向RNA干扰在削弱直肠癌免疫逃逸中的作用探讨

为探讨层黏素受体(laminin receptor,LNR)靶向RNA干扰在直肠癌免疫逃逸中的作用,将pGenesil-3-shLNR重组质粒转染SW480细胞,对比干扰组、对照组和空载组细胞中LNR mRNA和蛋白表达情况。建立SW480细胞与人Jurkat细胞Transwell小室旁分泌共培养模型,检测共培养48 h后SW480细胞和Jurkat细胞的凋亡率,并检测2种细胞Fas、FasL、Caspase-8 mRNA和蛋白表达情况。结果显示,重组质粒转染效率为(65.30±6.03)%。与对照组、空载组比较,干扰组LNR mRNA和蛋白相对表达量均较低(P<0.05);SW480细胞凋亡率较高(P<0.05),Jurkat细胞凋亡率较低(P<0.05);SW480细胞Fas、Caspase-8 mRNA和蛋白相对表达量均较高,FasL mRNA和蛋白相对表达量较低,Jurkat细胞Fas、Caspase-8 mRNA和蛋白相对表达量均较低,FasL mRNA和蛋白相对表达量较高(P<0.05)。对照组与空载组LNR mRNA及蛋白相对表达量,SW480细胞和Jurkat细胞凋亡率,SW480细胞和Jurkat细胞Fas、FasL、Caspase-8 mRNA及蛋白相对表达量比较,差异均无统计学意义(P> 0.05)。以上结果提示LNR靶向RNA干扰可抑制直肠癌细胞中LNR表达,可能通过调节SW480细胞和Jurkat细胞中Fas/FasL信号通路使SW480细胞诱导Jurkat细胞凋亡率降低,同时增强Jurkat细胞杀伤SW480细胞能力,削弱直肠癌细胞的免疫逃逸能力。

Laminin receptor targeting RNA interference diminishes immune escape of rectal cancer cells SW480

To investigate the role of laminin receptor(LNR) targeting RNA interference in immune escape of rectal cancer, pGenesil-3-shLNR recombinant plasmid was transfected into SW480 cells. The LNR mRNA and protein expressions of the interference group, the control group and the no-load group were compared. A trans-coculture model of SW480 cells and human Jurkat cells was established. After 48 hours’ coculture, apoptosis rates of SW480 cells and Jurkat cells were detected and Fas, FasL and Caspase-8 at both mRNA and protein levels were detected as well. The results showed that the efficiency of transfection of recombinant plasmid was(65.30±6.03)%. Compared with the control group and the no-load group, the relative expressions of LNR mRNA and protein in the interference group were reduced(P < 0.05), and the apoptosis rate of SW480 cells was higher(P < 0.05) and the apoptosis rate of Jurkat cells was lower(P < 0.05), the relative expressions of Fas and Caspase-8 mRNA and protein in SW480 cells were higher, the relative expressions of FasL mRNA and protein were lower, while the relative expressions of Fas and Caspase-8 mRNA and protein in Jurkat cells were higher and the relative expressions of FasL mRNA and protein were lower(P < 0.05). There were no significant differences in the relative expressions of LNR mRNA and protein, the apoptosis rates of SW480 cells and Jurkat cells, and the relative expressions of Fas, FasL and Caspase-8 mRNA and protein of SW480 cells and Jurkat cells between the control group and the no-load group(P > 0.05) In conclusion, LNR targeting RNA interference inhibits the expression of LNR in rectal cancer cells, reduces the apoptotic rate of Jurkat cells induced by SW480 cells by regulating Fas/FasL signaling pathway in SW480 cells and Jurkat cells, enhances the killing ability of Jurkat cells to SW480 cells, and attenuates the immune escape ability of rectal cancer cells.