HK3对结肠癌细胞免疫浸润、增殖和侵袭的影响及其机制研究

Effect of HK3 on immune invasion, proliferation and invasion of colon cancer cells

目的:探讨己糖激酶3(hexokinase-3,HK3)对结肠癌细胞RKO和HCT116增殖、周期、迁移及免疫浸润的影响。方法:使用生物信息学分析HK3在肿瘤和正常结肠样品的表达水平,探讨HK3与结肠癌免疫浸润的关系。将对照组(si-NC)和干扰HK3组(si-HK3)转染人结肠癌细胞RKO和HCT116,用CCK-8法、流式细胞术、划痕实验和Transwell小室实验分别检测干扰HK3对RKO和HCT116细胞增殖、周期和迁移的影响,同时通过Western blot实验检测各组细胞中上皮间质转化(epithelial mesenchymal transition,EMT)相关蛋白:波形蛋白(vimentin)和上皮钙黏蛋白(E-cadherin)的表达水平。结果:HK3与结肠癌中多种免疫细胞的浸润相关,且与多种免疫检查点具有共表达关系。和对照组(si-NC)相比,干扰HK3组(siHK3#1和si-HK3#2)HK3的蛋白表达水平显著降低(P<0.05);干扰HK3后RKO和HCT116细胞的增殖、迁移明显被抑制(P<0.05),且RKO和HCT116细胞的细胞周期被显著阻滞在S期(P<0.05)。同时,干扰HK3增加E-cadherin的表达,但抑制vimentin的表达(P<0.05)。结论:HK3影响结肠癌的免疫浸润水平,同时干扰HK3可通过抑制EMT行为抑制结肠癌细胞的细胞增殖、细胞周期和迁移能力。

Objective:To investigate the effects of hexokinase-3(HK3)on the proliferation,cell cycle,migration and immunological invasion of RKO and HCT116. Methods:The expression levels of HK3 in tumor and normal colon samples were analyzed by bioinformatics to evaluate the relationship between HK3 and immunological invasion of colon cancer. RKO and HCT116were transfected in the control group(si-NC)and the interference group(si-HK3)and then CCK-8,flow cytometry,scratch test and Transwell test were used to examine the effects of interference with HK3 on the proliferation,cell cycle and migration of RKO and HCT116 cells. The expression levels of vimentin and E-cadherin that are related to epithelial-mesenchymal transition(EMT)were detected by Western blot. Results:HK3 was found to be associated with infiltration of multiple immune cells and co-expression with multiple immune checkpoints in colorectal cancer. Compared with the control group(si-NC),the protein expression level of HK3 in the interference group(si-HK3#1 and si-HK3#2)was significantly decreased(P<0.05);the proliferation and migration of RKO and HCT116 cells were significantly inhibited(P<0.05);the cell cycle of RKO and HCT116 cells was arrested in S phase(P<0.05). In addition,the expression of E-cadherin protein was increased while that of vimentin protein was inhibited(P<0.05). Conclusion:HK3 affects the level of immunological invasion of colon cancer. Interference with HK3 inhibits the proliferation,cell cycle and migration of colon cancer cells by inhibiting EMT.