蒜素抑制人甲状腺乳头状癌TPC-1细胞生长的作用及机制

Effect and mechanism of Allicin inhibition on human papillary thyroid carcinoma TPC-1 cells

目的探讨蒜素(Allicin)抑制人甲状腺乳头状癌TPC-1细胞生长的作用及机制。方法将培养的TPC-1细胞分为对照组和不同浓度(10、20、30、40、50μg/mL)Allicin处理组。四甲基偶氮噻唑蓝(MTT)法测定不同浓度的Allicin处理24、48、72 h对TPC-1细胞生长的抑制作用。倒置显微镜和扫描电镜观察Allicin处理前后细胞形态和超微结构的变化。流式细胞术检测Allicin对细胞周期的影响。免疫细胞化学技术检测Allicin作用细胞后凋亡蛋白Bax和抗凋亡蛋白Bcl-2表达的变化。结果与对照组比较,不同浓度的Allicin均可抑制TPC-1细胞的生长,差异有统计学意义(P <0.05);Allicin对细胞的抑制率与时间和浓度两大要素相关。随着Allicin浓度的增加,G2/M期细胞所占比例明显升高,G0/G1期细胞所占比例降低,除对照组与10μg/mL Allicin组比较差异无统计学意义(P> 0.05),其他相邻浓度组两两比较差异均有统计学意义(P <0.05)。与对照组比较,各浓度Allicin处理组细胞内的Bcl-2表达量均降低,Bax表达量均升高,差异有统计学意义(P <0.05);各相邻浓度Allicin处理组Bcl-2及Bax表达量比较,差异均有统计学意义(P <0.05)。结论 Allicin具有抑制甲状腺乳头状癌TPC-1细胞生长的作用,其机制可能与Allicin抑制细胞增殖,下调Bcl-2表达,上调Bax表达从而诱导细胞凋亡有关。

Objective To investigate the effect and mechanism of Allicin inhibition on the growth of human thyroid papillary carcinoma TPC-1 cells.Methods The cultured TPC-1 cells were divided into the control group and the Allicin treatment groups with different concentrations(10,20,30,40,50μg/mL).Methylthiazoletrazolium(MTT)method was used to determine the inhibitory effect of different concentrations of Allicin treatment on the growth of TPC-1 cells at 24,48,72 h.Inverted microscope and scanning electron microscope were used to observe the change of cell morphology and ultrastructure after treatment by Allicin.Flow cytometry was used to test the effects of Allicin on cell cycle.Immunocytochemical technique was used to detect the effects on expression of intracellular apoptotic protein Bax and anti-apoptotic protein Bcl-2 after treatment by Allicin.Results Compared with the control group,different concentrations of Allicin could inhibit the growth of TPC-1 cells,the differences were statistically significant(P<0.05);the inhibition rate of Allicin on cells was related to time and concentration.With the increase of Allicin concentration,the proportion of G2/M phase cells was significantly increased,while that of G0/G1 phase cells was decreased.All pairwise comparison of adjacent concentration groups showed statistically significant differences(P<0.05)except the control group and 10μg/mL Allicin group(P >0.05).Compared with the control group,the intracellular Bcl-2 expression was decreased and Bax expression was increased in every concentration of Allicin treatment group,the differences were statistically significant(P<0.05);the expression levels of Bcl-2 and Bax in Allicin treatment groups with different adjacent concentrations were compared,the differences were statistically significant(P<0.05).Conclusion Allicin can inhibit the growth of TPC-1 cells of papillary thyroid cancer,and its mechanism may be related to inhibition of cell proliferation by Allicin and the induction of apoptosis by down-regulating Bcl-2 and up-regulating Bax.