蚕蛹复合氨基酸对人肝癌细胞SMMC-7721的抑制作用

Inhibition Effect of Composite Amino Acid of Silkworm Chrysalis on Human Hepatic Cancer Cells SMMC-7721

本实验探讨蚕蛹复合氨基酸对人肝癌细胞株SMMC-7721的抑制作用。实验经MTT法检测药物对人正常肝脏细胞株QSG-7701的毒性后,计算药物安全浓度。将不同浓度的蚕蛹复合氨基酸与人肝癌细胞SMMC-7721共培养,采用MTT法测定OD值,评定蚕蛹蛋白复合氨基酸对肝癌细胞株的增殖抑制作用;经Ho-echst33258染色和倒置显微镜进行形态学观察以检测细胞凋亡率;流式细胞法测定细胞周期和Annexin V/PI双染色法检测细胞凋亡。细胞毒性实验表明:蚕蛹蛋白复合氨基酸的最大无毒浓度为10 mg/mL。不同浓度的蚕蛹蛋白复合氨基酸对SMMC-7721细胞均有抑制作用,各组与对照组相比差异有显著性(P<0.05),并呈剂量和时间依赖性。Hoechst33258染色和流式细胞术结果亦证实,蚕蛹复合氨基酸能显著促进SMMC-7721细胞的凋亡。

The objective of this study was to investigate the inhibition effect of composite amino acid of silkworm chrysalis （CAASCP） on human hepatic cancer cells SMMC-7721. After the detection of safe concentration of the drug using cell line QSG-7701 by M33＂ assays, the inhibition effects of CAASCP against hepatic cancer cells SMMC-7721 were evaluated with different concentrations of CAASCP using MTr methods; the ratios of apoptosis were tested with morphology under inverted microscope using Hoechst33258 dyeing;cell cycles were determined by flow cytometry with PI staining. Cell apoptosis was also detected with Annexin V/PI double staining using flow cytometry. The results showed that the TC0 of CAASCP was 10 mg/mL in cytotoxicity experiments. All concentrations of CAASCP had inhibited the proliferation of cell line SMMC-7721 significantly,compared with negative control group （P 〈 0.05 ）, and presented in dose and time-dependent manner. The Hoechst33258 dyeing and flow cytometry tests also proved that CAASCP can promote apoptosis of SMMC-7721 cell strain obviously.