融合受体eDR4/iFas提高sTRAIL对HT-29细胞凋亡的敏感性

中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (9): 966-972.

融合受体eDR4/iFas提高sTRAIL对HT-29细胞凋亡的敏感性

李招发^{1, 2}, 王从阳¹, 邓小英¹, 惠二京¹

¹ 华侨大学生物医学学院,泉州 362021,福建;
² 华侨大学分子药物教育部工程研究中心,泉州 362021,福建

收稿日期:2013-09-23 修回日期:2014-06-26 出版日期:2014-09-26 发布日期:2014-09-26
作者简介:李招发,男,博士,副研究员,研究方向:基因工程与基因治疗。 Tel: 0595-22690838 E-mail: lizhaofa@hqu.edu.cn
基金资助:
中央高校基本科研业务费资助项目 (JB-ZR1142); 福建省自然科学基金 (2010J01207)资助

Infusion receptor eDR4/iFas enhances TRAIL-induced apoptosis of HT-29 cells

LI Zhao-fa^{1, 2}, WANG Cong-yang¹, DENG Xiao-ying¹, HUI Er-jing¹

¹ School of Biomedical Sciences, Huaqiao University, Quanzhou 362021,Fujian, China;
² Molecular Medicine Engineering Research Center of the Ministry of Education, Huaqiao University, Quanzhou 362021, Fujian, China

Received:2013-09-23 Revised:2014-06-26 Online:2014-09-26 Published:2014-09-26

摘要/Abstract

摘要： 目的探讨sTRAIL联合融合受体eDR4/iFas对HT-29细胞的凋亡作用。方法采用PCR方法分别扩增eDR4和iFas,通过重叠PCR将两者连接起来组成融合受体基因eDR4/iFas,将其克隆到带Survivin启动子的表达载体上, 另外采用组成型启动子CAG调控sTRAIL表达。体外转染结直肠癌细胞株HT-29和人胚肾细胞HEK293,MTT法检测两种细胞的存活率,RT-PCR和Western Blot检测目的蛋白的表达, Hoechst染色进一步分析eDR4/iFas联合sTRAIL对HT-29细胞的凋亡效果。结果在HT-29细胞中,sTRAIL组的细胞存活率为 40.9%±18.7%,而eDR4/iFas+sTRAIL组的细胞存活率为 21.6%±9.1%,两者间存在统计学差异(P<0.05)。结论: eDR4/iFas能提高sTRAIL对HT?29细胞凋亡的敏感性,而对正常细胞HEK293没有毒副作用。

关键词: sTRAIL, 融合受体, eDR4/iFas, Survivin启动子, 结直肠癌细胞株HT-29

Abstract: AIM: To investigate the effect of sTRAIL combined with infusion receptor eDR4/iFas on apoptosis of HT-29 cells. METHODS: In this study, eDR4 and iFas genes were amplified respectively by PCR and used to generate fusion gene named eDR4/iFas by splicing overlap extension PCR. eDR4/iFas was cloned into the expression vector with the tumor-specific survivin promoter, and expressed sTRAIL with constitutive promoter CAG. After treatment with eDR4/iFas and sTRAIL, the cell viability analysis of both colorectal cancer cells HT-29 and human embryonic kidney cells HEK293 was assayed by MTT. Both TRAIL mRNA and protein expressions were detected with RT-PCR and ELISA respectively. The apoptosis of HT-29 cells was analyzed by Hoechst staining. RESULTS: In HT-29 cells, cell survival rate in sTRAIL group was 40.9%±18.7%, while cell survival rate in eDR4/iFas+sTRAIL group was 21.6%±9.1%, there was significant difference between the two groups (P<0.05). CONCLUSION: The results have revealed that eDR4/iFas can improve TRAIL-induced apoptosis of HT-29 cells, has no toxicity for HEK293 cells.

Key words: sTRAIL, infusion receptor, eDR4/iFas, survivin promoter, colorectal cancer cell line HT-29

中图分类号:

R73-36⁺2

李招发, 王从阳, 邓小英, 惠二京. 融合受体eDR4/iFas提高sTRAIL对HT-29细胞凋亡的敏感性[J]. 中国临床药理学与治疗学, 2014, 19(9): 966-972.

LI Zhao-fa, WANG Cong-yang, DENG Xiao-ying, HUI Er-jing. Infusion receptor eDR4/iFas enhances TRAIL-induced apoptosis of HT-29 cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(9): 966-972.

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