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目的研究丹参酮ⅡA(TanⅡA)对HL-60细胞的人端粒酶反转录酶(hTERT)表达的影响,探讨TanⅡ诱导肿瘤细胞凋亡的分子机制。方法用RPMI 1640培养HL-60细胞,培养24 h后分为3组[TanⅡA组、全反式维A酸(ATRA)组、对照组],分别加入500μg.L-1TanⅡA、500μg.L-1ATRA和0.1 mL.L-1二甲基亚砜治疗5 d。加药前和加药后每天分别收集细胞,采用锥虫蓝染色后计数活细胞,流式细胞仪(FCM)检测HL-60细胞凋亡率,半定量RT-PCR法测定HL-60细胞hTERT相对表达水平。结果与对照组比较,药物治疗2 d后,TanⅡA组和ATRA组HL-60细胞的生长均明显受到抑制(Pa<0.05),TanⅡA组和ATRA组的抑制作用比较差异无统计学意义(P>0.05)。TanⅡA组和ATRA组在药物治疗2 d后细胞凋亡率分别为32.11%,23.31%,均明显高于对照组(6.08%)(Pa<0.05),且细胞凋亡率逐日上升,而对照组细胞凋亡率无明显变化。药物处理1 d后,TanⅡA组和ATRA组的HL-60细胞hTERT相对表达水平均明显低于对照组(Pa<0.05),且表达水平逐日下降,而对照组表达水平无明显变化。结论丹参酮ⅡA抑制HL-60细胞生长、诱导细胞凋亡可能与下调hTERT基因表达水平有关。
Objective To study the effect of TanⅡA on the expression of human telomerase reverse transcriptase (hTERT) in HL-60 cells and to explore the molecular mechanism of TanⅡ-induced tumor cell apoptosis. Methods HL-60 cells were cultured in RPMI 1640 and divided into 3 groups [TanⅡA group, ATRA group and control group] after 24 h culture, and were respectively treated with 500 μg.L-1TanⅡA and 500 μg.L-1 ATRA And 0.1 mL.L-1 dimethyl sulfoxide for 5 d. The cells were collected daily before dosing and after dosing. The viability of HL-60 cells was measured by flow cytometry (FCM). The number of hTERT cells in HL-60 cells was measured by semi-quantitative RT-PCR Relative expression level. Results Compared with the control group, the growth of HL-60 cells in TanⅡA group and ATRA group was significantly inhibited (Pa> 0.05) after 2 days of drug treatment, while there was no significant difference in inhibition between TanⅡA group and ATRA group (P> 0.05). The apoptosis rates of TanⅡA group and ATRA group after treatment for 2 d were 32.11% and 23.31%, respectively, which were significantly higher than those in control group (6.08%, Pa <0.05) There was no significant change in apoptosis rate. After 1 day of drug treatment, the relative expression level of hTERT in HL-60 cells in Tan II A group and ATRA group was significantly lower than that in control group (Pa <0.05), and the expression level decreased day by day, while the expression level in control group did not change significantly. Conclusion Tanshinone Ⅱ A can inhibit the growth of HL-60 cells and induce the apoptosis of HL-60 cells, which may be related to the downregulation of hTERT gene expression.