腺病毒介导的人环氧化酶-2反义RNA对肝癌生长的影响

Effects of adenovirus-mediated human cyclooxygenase-2 antisense RNA on the growth of hepatocellular carcinoma.

作者信息

Wang Xiao-Hu, Li Sheng-Bao, Tong Qiang, Xie Guo-Jian, Wu Qing-Ming

机构信息

Department of Gastroenterology, Taihe Hospital, Yunyang Medical College, Shiyan 442000, Hubei Province, China.

出版信息

World J Gastroenterol. 2005 Oct 21;11(39):6110-4. doi: 10.3748/wjg.v11.i39.6110.

DOI:10.3748/wjg.v11.i39.6110

PMID:16273635

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4436396/

Abstract

AIM

To investigate the relation between the expression of cyclooxygenase-2 (COX-2) and liver cancer, to construct the recombinant adenovirus encoding human COX-2 antisense RNA, and to explore its effects on liver cancer cell proliferation.

METHODS

We studied the expression of COX-2 in 34 cases of hepatocellular carcinoma (HCC) and SMMC7402 and SMMC7721 by immunohistochemical technique. Recombinant adenovirus Ad-AShcox-2 was constructed and transfected into human HCC cell lines SMMC7402 and SMMC7721, and its effects on COX-2 expression, cell apoptosis and cell cycle were analyzed by flow cytometry. Cell proliferation was determined by colony-forming efficiency.

RESULTS

We observed COX-2 expression in 82.4% of the HCC and SMMC7402 cells, but no COX-2 expression in SMMC7721 cells. In addition, recombinant adenovirus encoding antisense COX-2 fragment Ad-AShcox-2 was obtained with the titer of 1.06 x 10(12) PFU/mL. Ad-AShcox-2 could reduce the expression of COX-2 and enhance the percentage of cells in G(1)/G(0) phase in SMMC7402 cell line. The difference of apoptotic index between the Ad-AShcox-2 group and control group was statistically significant (t( control group ) = 32.62 and t( Ad-LacZ ) = 10.93, P<0.001) in SMMC7402 but not in SMMC7721. Similarly, colony-forming rates of SMMC7402 and SMMC7721 cell lines, after the transfer of Ad-AShcox-2, were (2.7+/-0.94)% and (33.6+/-4.24)%, respectively.

CONCLUSION

Reduction in the expression of COX-2 can inhibit COX-2 expressing HCC cells.

摘要

目的

研究环氧合酶-2（COX-2）表达与肝癌的关系，构建编码人COX-2反义RNA的重组腺病毒，并探讨其对肝癌细胞增殖的影响。

方法

采用免疫组织化学技术研究34例肝细胞癌（HCC）及SMMC7402和SMMC7721细胞中COX-2的表达。构建重组腺病毒Ad-AShcox-2并转染人肝癌细胞系SMMC7402和SMMC7721，通过流式细胞术分析其对COX-2表达、细胞凋亡和细胞周期的影响。通过集落形成效率测定细胞增殖。

结果

我们观察到82.4%的HCC和SMMC7402细胞中有COX-2表达，但SMMC7721细胞中无COX-2表达。此外，获得了编码反义COX-2片段的重组腺病毒Ad-AShcox-2，滴度为1.06×10(12) PFU/mL。Ad-AShcox-2可降低SMMC7402细胞系中COX-2的表达，并增加G(1)/G(0)期细胞百分比。在SMMC7402中，Ad-AShcox-2组与对照组的凋亡指数差异有统计学意义（对照组t = 32.62，Ad-LacZ组t = 10.93，P<0.001），而在SMMC7721中无差异。同样，转染Ad-AShcox-2后，SMMC7402和SMMC7721细胞系的集落形成率分别为(2.7±0.94)%和(33.6±4.24)%。