长春瑞滨对人肺腺癌细胞体外凋亡及端粒酶活性表达的影响

[Effects of vinorelbine on apoptosis and expression of telomerase activity in human lung adenocarcinoma cells in vitro].

作者信息

Liu Jing-jing, Chen Gong-yan, Wang Meng, Yang Zhao-yang, Hong Xuan

机构信息

Tumor Hospital of Harbin Medical University, Harbin 150040, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2010 Oct;32(10):743-7.

PMID:21163063

Abstract

OBJECTIVE

To study the effect of vinorelbine on apoptosis, telomerase activity and expression of hTERT gene in human lung adenocarcinoma cell line Anip973 cells.

METHODS

Anip973 cells were cocultured with Vinorelbine at different concentrations and collected at 24 h, 48 h and 72 h after treatment, respectively. The inhibition rate of cell growth was determined by methyl thiazolyl tetrazolium (MTT) assay to evaluate the effect of vinorelbine. The percentage of apoptosis was detected by flow cytometry. The cellular morphology was observed under inverted microscope and electron microscope. Telomerase activity of Anip973 cells was determined by the method of TRAP-PAGE-silver staining. RT-PCR was used to detect the expression of hTERT mRNA.

RESULTS

Vinorelbine down-regulated the telomerase activity and expression of hTERT gene mRNA, inhibited the growth of Anip973 cells, and induced cell apoptosis in a time- and concentration-dependent manner. Annexin V-FITC assay showed 0.08 µg/ml NVB group could inhibited cell proliferation in 24 hour, and apoptosis rate was (7.37 ± 0.35)%, RT-PCR detection of hTERT mRNA expression in this group was (57.01 ± 1.71), and they were very significantly different from that in the control group (P < 0.01), but telomerase activity was (6.36 ± 0.06), compared with the control group showed no significant difference(P > 0.05). The change of hTERT mRNA expression is more sensitive than telomerase activity. The apoptosis rate, telomerase activity and hTERT mRNA expression of 0.4 µg/ml group and 2.0 µg/ml group were very significantly different from that in the control group (P < 0.01). The telomerase activity of 2.0 µg/ml at 72 hour group was (1.36 ± 0.27), basically completely inhibited, while the apoptosis rate was (74.87 ± 1.88)%, showed the cell apoptosis rate was in a negative correlation with the down-regulation of the hTERT mRNA (r = -0.96046, P < 0.01).

CONCLUSIONS

Vinorelbine can induce apoptosis in Anip973 cells, and its mechanism of action is related to telomerase activity. The detection of telomerase activity and expression of hTERT gene is useful in predicting prognosis of patients with lung adenocarcinoma.

摘要

目的

研究长春瑞滨对人肺腺癌细胞系Anip973细胞凋亡、端粒酶活性及hTERT基因表达的影响。

方法

将Anip973细胞与不同浓度长春瑞滨共培养，分别于处理后24 h、48 h和72 h收集细胞。采用甲基噻唑基四氮唑（MTT）法测定细胞生长抑制率以评估长春瑞滨的作用效果。通过流式细胞术检测凋亡率。在倒置显微镜和电子显微镜下观察细胞形态。采用TRAP-PAGE-银染法测定Anip973细胞的端粒酶活性。用RT-PCR检测hTERT mRNA的表达。

结果

长春瑞滨下调端粒酶活性及hTERT基因mRNA表达，抑制Anip973细胞生长，并呈时间和浓度依赖性诱导细胞凋亡。Annexin V-FITC检测显示，0.08 μg/ml长春瑞滨组在24 h可抑制细胞增殖，凋亡率为（7.37±0.35）%，该组RT-PCR检测hTERT mRNA表达为（57.01±1.71），与对照组相比差异极显著（P<0.01），但端粒酶活性为（6.36±0.06），与对照组相比无显著差异（P>0.05）。hTERT mRNA表达变化比端粒酶活性更敏感。0.4 μg/ml组和2.0 μg/ml组的凋亡率、端粒酶活性及hTERT mRNA表达与对照组相比差异极显著（P<0.01）。2.0 μg/ml 72 h组端粒酶活性为（1.36±0.27），基本完全被抑制，而凋亡率为（74.87±1.88）%，表明细胞凋亡率与hTERT mRNA下调呈负相关（r=-0.96046，P<0.01）。