抑制LILRB2对人结直肠癌SW480细胞增殖和凋亡的影响

Effect of inhibiting LILRB2 on proliferation and apoptosis of human colorectal cancer SW480 cells

目的探索LILRB2对结直肠癌SW480细胞增殖和凋亡的影响,并进一步探索其机制。方法体外培养结直肠癌SW480细胞,并将其分为空白对照组、阴性对照组及实验组。利用流式细胞仪检测LILRB2的表达情况。利用qPCR检测LILRB2的表达,将空载体质粒和LILRB2质粒分别转染入SW480细胞;利用CCK-8法检测细胞增殖情况;流式细胞术检测细胞凋亡;Western blot实验对蛋白表达变化进行检测。结果LILRB2在SW480中的表达量为0.84±0.09,较FHC细胞提升两倍0.38±0.05,差异具有统计学意义(P<0.05)。病毒感染后实验组SW480细胞中LILRB2的表达量(0.48±0.07)明显降低。CCK-8实验结果所示,处理12 h后LILRB2低表达实验组的SW480细胞增殖情况被抑制,LILRB2低表达的实验组的SW480细胞细胞凋亡比例上升为49.3%±1.2%,与空白对照组和阴性对照组的细胞凋亡比例(7.48%±0.85%)、(7.35%±0.93%)相比,差异具有统计学意义(P<0.05)。LILRB2低表达的实验组SW480细胞的活性氧水平(3.34±0.29)远高于与空白对照组(1.03±0.12)和阴性对照组(1.07±0.09),差异有统计学意义(P<0.05);在加入ROS清除剂NAC之后,LILRB2低表达实验组细胞凋亡上升。结论LILRB2低表达抑制SW480细胞的增殖,诱导细胞凋亡,其可能通过调控ROS水平发挥作用,为LILRB2在结直肠癌的研究中提供一定的理论基础。

Objective To explore the effect of LILRB2 on the proliferation and apoptosis of colorectal cancer SW480 cells,and to further explore its mechanism.Methods Colorectal cancer SW480 cells were cultured in vitro and divided into blank control group,negative control group and experimental group.The expression of LILRB2 was detected by flow cytometry.The expression of LILRB2 was detected by qPCR,and the empty vector plasmid and the LILRB2 plasmid were transfected into SW480 cells respectively;cell proliferation was detected by CCK-8 method;cell apoptosis was detected by flow cytometry.Western blot was used to detect changes in the expression of related proteins.Results The expression level of LILRB2 in SW480 was 0.84±0.09,twice higher than that in FHC cells(0.38±0.05),and the difference was statistically significant(P<0.05).After virus infection,the expression of LILRB2(0.48±0.07)in SW480 cells of the experimental group decreased significantly.CCK-8 experiment results showed that after 12 hours of treatment,the proliferation of SW480 cells in the LILRB2 low expression experimental group was inhibited,and the percentage of apoptosis in SW480 cells in the LILRB2 low expression experimental group increased to 49.3%±1.2%,which was statistically significant(P<0.05)compared with the percentage of apoptosis in the blank control group and the negative control group(7.48%±0.85%,7.35%±0.93%).The ROS level of SW480 cells in the experimental group with low LILRB2 expression was significantly higher than that in the blank control group and negative control group(P<0.05).After adding ROS scavenger NAC,the apoptosis of LILRB2 in the experimental group increased.Conclusion The low expression of LILRB2 inhibits the proliferation of SW480 cells and induces apoptosis,which may play a role by regulating the level of ROS,providing a theoretical basis for the study of LILRB2 in colorectal cancer.