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IARS2基因在结肠癌中的表达及其敲低对RKO细胞生物学行为的影响。

Expression of IARS2 gene in colon cancer and effect of its knockdown on biological behavior of RKO cells.

作者信息

Zhong Ling, Zhang Yi, Yang Jing-Yu, Xiong Liang-Fa, Shen Tao, Sa Ya-Lian, O'Yang Yi-Ming, Zhao Si-Hui, Chen Jia-Yong

机构信息

Kunming Medical University Chenggong District, Kunming 650500, Yunnan Province, China ; Department of Emergency Intensive Care Unit, The First People's Hospital of Yunnan Province Kunming 650032, Yunnan Province, China.

Department of Thyroid and Breast Surgery, The Second Affiliated Hospital of Kunming Medical University Kunming 650101, Yunnan Province, China.

出版信息

Int J Clin Exp Pathol. 2015 Oct 1;8(10):12151-9. eCollection 2015.

Abstract

OBJECTIVE

To investigate the expression level of IARS2 gene in colon cancer tissues and various cell strains of the cancer; to explore cytologically the effect of IARS2 gene knockdown on proliferation, apoptosis and cell cycle of RKO cells in the cancer.

METHODS

Real-time, fluorescence-based quantitative PCR (qPCR) was used to detect the expression of IARS2 gene in human colon cancer and surrounding tissues and in various cell strains of the cancer; the RNA interference target of IARS2 gene was designed and the target was detected by Western blot; the IARS2-siRNA lentiviral vector was established and used to infect the RKO cells of colon cancer; qPCR was employed to determine the effect of gene knockdown; changes of the RKO cells in growth, apoptosis, cell cycle and clone formation were observed after IARS2 gene knockdown.

RESULTS

The expression of IARS2 gene was higher in human colon cancer tissues than in surrounding tissues; there was expression of IARS2 gene in colon cancer cells, and the expression level of IARS2 gene mRNA was higher in the RKO cells than in the SW480, HCT116, DLD1, HT-29 and SW620 cells. After infection of the RKO cells with IARS2-siRNA lentivirus, the expression of IARS2 gene was inhibited in the level of mRNA; proliferation rate of the RKO cells was significantly inhibited; the G1 phase arrest of the RKO cells was increased with less RKO cells in S phase; the apoptotic RKO cells increased significantly; and the number of colonies of the RKO cells reduced.

CONCLUSION

The expression of IARS2 gene is different in human colon cancer and surrounding tissues; after knockdown of IARS2 gene, proliferation of the RKO cells is inhibited; there are more cells in G phase and fewer cells in S phase; apoptosis of cells is increased; and formation of colonies is reduced. IARS2 gene is probably a cancer-promoting gene.

摘要

目的

研究IARS2基因在结肠癌组织及该癌症的各种细胞株中的表达水平;从细胞学角度探讨IARS2基因敲低对结肠癌RKO细胞增殖、凋亡及细胞周期的影响。

方法

采用实时荧光定量PCR(qPCR)检测IARS2基因在人结肠癌组织、癌旁组织及该癌症的各种细胞株中的表达;设计IARS2基因的RNA干扰靶点并通过蛋白质免疫印迹法进行检测;构建IARS2-siRNA慢病毒载体并用于感染结肠癌RKO细胞;采用qPCR确定基因敲低的效果;观察IARS2基因敲低后RKO细胞在生长、凋亡、细胞周期及克隆形成方面的变化。

结果

IARS2基因在人结肠癌组织中的表达高于癌旁组织;在结肠癌细胞中有IARS2基因表达,且IARS2基因mRNA的表达水平在RKO细胞中高于SW480、HCT116、DLD1、HT-29和SW620细胞。用IARS2-siRNA慢病毒感染RKO细胞后,IARS2基因的mRNA水平表达受到抑制;RKO细胞的增殖率显著受到抑制;RKO细胞的G1期阻滞增加,S期细胞减少;凋亡的RKO细胞显著增加;RKO细胞的集落数减少。

结论

IARS2基因在人结肠癌组织与癌旁组织中的表达存在差异;IARS2基因敲低后,RKO细胞的增殖受到抑制;G期细胞增多,S期细胞减少;细胞凋亡增加;集落形成减少。IARS2基因可能是一个促癌基因。

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