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[组蛋白去乙酰化酶6下调对人喉癌细胞系Hep-2裸鼠移植瘤生长的影响及机制研究]

[Effect of HDAC6 down-regulation on the growth of xenografted human laryngeal carcinoma cell line Hep-2 in nude mice and underlying mechanism].

作者信息

Yuan Lin-lin, Lou Wei-hua, Sang Jian-zhong

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, the First Affiliated Hospital of Zhengzhou University, China.

出版信息

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2012 Jun;47(6):481-6.

Abstract

OBJECTIVE

To study the effect of histone deacetylation 6 (HDAC6) siRNA on the growth of xenografted human laryngeal squamous cell carcinoma cell line Hep-2 in nude mice and underlying mechanism.

METHODS

Laryngeal squamous cell carcinoma cell line Hep-2 cells were subcutaneously injected to the back of nude mice and transplanted tumor model was established after one week. Nude mice was divided into three groups including blank control group, empty vector group and HDAC6 siRNA group, and the tumor growth was observed. Ki-67 proliferation index was detected by immunohistochemistry. Western blot, in situ hybridization and immunohistochemistry were used to detect the mRNA and protein expressions of HDAC6 in xenograft. The expressions of Bcl-2 and Bax proteins were examined by Western blotting. Cell apoptosis was detected by TUNEL.

RESULTS

The mean volume of xenograft transfected with HDAC6 siRNA was less than that of xenograft transfected with empty vector or that of xenograft with blank control treatment (P < 0.05). HDAC6 siRNA effectively down-regulated the expressions of HDAC6 mRNA and the expressions of HDAC6 and Bcl-2 proteins, but up-regulated the expression of Bcl-2 protein in xenografts, with significant differences (all P < 0.05). The proliferation index of Ki-67 in HDAC6 siRNA transfection group was significantly lower than that in blank control group or empty vector group (P < 0.05). TUNEL assay demonstrated that HDAC6 evidently evoked cell apoptosis (P < 0.05).

CONCLUSION

HDAC6 siRNA could effectively inhibited the growth of xenografted human laryngeal carcinoma cell line Hep-2 in nude mice, down-regulate the expressions of HDAC6 and bcl-2, and up-regulate the expression of bax.

摘要

目的

研究组蛋白去乙酰化酶6(HDAC6)小干扰RNA(siRNA)对人喉鳞状细胞癌细胞系Hep-2裸鼠移植瘤生长的影响及其潜在机制。

方法

将喉鳞状细胞癌细胞系Hep-2细胞皮下注射到裸鼠背部,1周后建立移植瘤模型。将裸鼠分为空白对照组、空载体组和HDAC6 siRNA组,观察肿瘤生长情况。采用免疫组织化学法检测Ki-67增殖指数。运用蛋白质印迹法、原位杂交法和免疫组织化学法检测移植瘤中HDAC6的mRNA和蛋白表达。通过蛋白质印迹法检测Bcl-2和Bax蛋白的表达。采用TUNEL法检测细胞凋亡。

结果

HDAC6 siRNA转染的移植瘤平均体积小于空载体转染的移植瘤及空白对照处理的移植瘤(P<0.05)。HDAC6 siRNA有效下调了移植瘤中HDAC6 mRNA的表达以及HDAC6和Bcl-2蛋白的表达,但上调了Bax蛋白的表达,差异均有统计学意义(均P<0.05)。HDAC6 siRNA转染组的Ki-67增殖指数明显低于空白对照组和空载体组(P<0.05)。TUNEL检测显示HDAC6明显诱导细胞凋亡(P<0.05)。

结论

HDAC6 siRNA可有效抑制人喉癌细胞系Hep-2裸鼠移植瘤的生长,下调HDAC6和bcl-2的表达,并上调bax的表达。

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