低表达人血细胞特异蛋白1相关蛋白X1对人结肠癌细胞增殖的影响

Effect of low expression of human hemocyte specific protein 1 associated protein X1 on proliferation of human colon cancer cells

目的观察低表达人血细胞特异蛋白1相关蛋白X1(HAX1)对结肠癌细胞增殖的影响。方法利用HAX1特异性低表达小干扰RNA(siHAX1)瞬时转染结肠癌细胞SW48,通过Western blot验证低表达成功,采用细胞计数试剂盒-8(CCK-8)、克隆形成实验观察HAX1对SW48细胞的影响,利用Western blot检测细胞周期蛋白(Cyclin) D1表达。结果Western blot证实在结肠癌细胞中低表达HAX1模型的成功建立,CCK-8实验结果表明,对照组(siCon)48 h结肠癌细胞吸光度(A)值(1.264±0.020)较各实验组(siHAX1)A值(1.122±0.057、1.021±0.033、0.989±0.049)差异有统计学意义(P<0.05),即实验组增殖能力降低。72 h时,对照组A值(1.332±0.019),各实验组A值(1.199±0.040、1.081±0.022、1.068±0.026),组间比较差异有统计学意义(P<0.05),说明72 h时实验组增殖能力仍降低。克隆形成实验显示,对照组(siCon)克隆形成数目[(713±40)个]明显多于各实验组克隆形成数目[(422±33)、(558±37)、(505±23)个],且差异有统计学意义(P<0.05),说明实验组克隆形成能力降低。Western blot证实在细胞低表达HAX1蛋白后,可明显降低Cyclin D1蛋白的表达。结论在结肠癌细胞中,低表达HAX1可通过抑制Cyclin D1的表达抑制肿瘤细胞增殖。

Objective To observe the effect of human hemocyte specific protein 1 (HS-1) associated protein X-1on the proliferation of colon cancer cells. Methods HAX1-specific small interfering RNA (siHAX1) was transiently transfected into colon cancer cells SW48, which was verified by the western blotting. The effect of HAX1 on SW48 cells was observed by cell counting kit-8 (CCK-8) and clone formation assay. At the same time, the expression of Cyclin D1 was showed by Western blotting. Results It was confirmed that the expression of HAX1 was declined in colon cancer cells by western blotting. And, at the 48 h, the A value (1.264±0.020) of the colon cancer cells in the control group (siCon) was higher than that in the experimental group (siHAX1)(1.122±0.057, 1.021±0.033, 0.989±0.049), which was statistically significant (P<0.05). In other words, the proliferation of experimental group was decreased. At the 72 h, the A value (1.332±0.019) in the control group was different from that in the experimental group (1.199±0.040, 1.081±0.022, 1.068±0.026), and the difference was statistically significant (P<0.05). So the proliferative capacity of the experimental group was still decreased. In addition, the expression of Cyclin D1 protein was significantly decreased after low expression of HAX1 protein. Conclusion Low expression of HAX1 in colon cancer cells can inhibit the proliferation of cells by decreasing the expression of Cyclin D1.