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- 2017
柴胡皂甙d对人肝癌细胞BECN1表达及自噬功能的影响
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Abstract:
摘要：目的 观察柴胡皂甙d(SSd)对肝癌细胞SMMC-7721生长增殖及自噬作用的影响，初步探讨其可能的机制。方法 肝癌SMMC-7721细胞株经不同终浓度SSd(5.0、7.5、10.0、12.5、15.0、和17.5mg/L)作用24、48、72h，MTT法检测细胞增殖，筛选最适作用浓度及时间，RT-PCR测定自噬基因BECN1 mRNA表达水平，Western blot检测BECN1蛋白表达水平。结果 SMMC-7721细胞增殖的抑制率随着SSd浓度的增大而增大，浓度达12.5mg/L时，抑制率最高(60%)；SSd作用后BECN1在基因和蛋白水平的相对表达量比对照组明显升高(P<0.05)，3-甲基腺嘌呤(3-MA)可降低BECN1基因和蛋白的表达，并可降低SSd作用后的BECN1的表达。结论 SSd对肝癌细胞SMMC-7721增殖的抑制作用呈现浓度和时间的依赖性，基本符合药物剂量-效应关系。SSd可能通过上调自噬基因BECN1的表达诱导自噬性死亡的发生，从而达到抑制肝癌细胞SMMC-7721增殖的作用。
ABSTRACT: Objective To observe the influence of saikosaponin-d (SSd) on the proliferation and the function of autophagy of human hepatocellular carcinoma (HCC) cell line SMMC-7721 to explore the possible mechanisms. Methods SMMC-7721 was cultured in vitro and then treated with SSd of various concentrations (5.0, 7.5, 10.0, 12.5, 15.0 and 17.5mg/L) for 24, 48 and 72h. We used MTT to detect cell proliferation, selected the optimal concentration and time, and detected the expressions of BECN1 at mRNA and protein levels by PCR and Western blot. Results The inhibition rate of the proliferation of SMMC-7721 cell line increased with the increase of the concentration of SSd, and the highest inhibition rate (60%) appeared when the concentration reached 12.5mg/L. The expression of BECN1 in the group with SSd was obviously higher than that in the control group (P<0.05). 3-MA decreased not only the expressions of BECN1 at mRNA and protein levels but also the expression of BECN1 when used in conjunction with SSd. Conclusion The inhibiting function of SSd on SMMC-7721 presents a dependency between drug concentration and function time, basically in line with the drug dose-effect relationship. SSd induces the occurrence of autophagic cell death through up-regulating the expression of BECN1, thus inhibiting the proliferation of SMMC-7721
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