壁虎粗多肽对肝癌HepG2细胞增殖及自噬相关蛋白表达的影响

Effect of gecko crude poly peptide on the proliferation- and autophagy- related protein expression of hepatocellular carcinoma HepG2 cells

目的观察壁虎粗多肽对肝癌HepG2细胞增殖及自噬相关蛋白表达的影响。方法低、中、高剂量实验组肝癌HepG2细胞分别予以0.15,0.30,0.45mg·mL-1壁虎粗多肽处理,对照组细胞则予以等量0.9%氯化钠。噻唑蓝(MTT)法检测HepG2细胞增殖抑制情况,用蛋白免疫印迹法检测自噬相关蛋白表达情况。结果给药72h后,低、中、高剂量实验组的增殖抑制率分别为(23.16±2.17)%,(51.24±2.28)%,(89.59±2.94)%,组间比较差异均有统计学意义(均P<0.05)。对照组及低、中、高剂量实验组的微管轻链Ⅰ蛋白3-Ⅱ型(LC3Ⅱ)相对表达量分别为0.43±0.12,0.45±0.11,0.76±0.15,0.87±0.16;Beclin-1蛋白表达量分别为0.41±0.09,0.46±0.12,0.48±0.14,0.91±0.15,中、高剂量实验组的LC3Ⅱ和Beclin-1蛋白分别与对照组比较,差异均有统计学意义(均P<0.05)。结论壁虎粗多肽可有效抑制肝癌HepG2细胞增殖,其作用机制可能与上调自噬相关蛋白表达相关。

Objective To observe the effect of gecko crude polypeptide on the proliferation and autophagy-related protein expression of hepatocellular carcinoma HepG2 cells. Methods The hepatocellular carcinoma HepG2 cells in experimental-L/-M/-H groups were treated by 0.15, 0.30, 0.45 mg·mL-1 gecko crude peptides, respectively, while the cells in control group were treated with equal of 0.9%NaCl. The proliferation inhibition of HepG2 cells were detected by Methyl thiazolyl tetrazolium(MTT);the autophagy related protein expression were observed and detected by western blot. Results There were statistically significant differences among the experimental-L/-M/-H groups in the proliferation inhibition rates of HepG2 cells [(23.16±2.17)% vs(51.24±2.28)% vs(89.59±2.94)%](P<0.05) at 72 h after treatment. The relative expression of microtubule associated proteinⅠ 1 ight chain3-Ⅱ type(LC3Ⅱ) in control group and experimental-L/-M/-H groups were 0.43±0.12, 0.45±0.11, 0.76±0.15, 0.87±0.16, respectively;the Beclin-1 protein relative expression were 0.41±0.09, 0.46±0.12, 0.48±0.14, 0.91±0.15, respectively. The differences between the control group and the experimental-M/-H group in the expression of LC3Ⅱ and Beclin-1 protein of was statistically significant( P < 0. 05). Conclusion Gecko crude peptides can inhibit the proliferation of hepatocellular carcinoma HepG2 cells by up-regulating the expression of autophagy related proteins.