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采用实时逆转录聚合酶链反应(Real-time RT-PCR)方法,研究A、C型荚膜合成酶与抗癌序列的基因融合对诱导凋亡相关基因BCL-2、BAX和Caspase-3表达的影响。

Studying the effect of gene fusion of A and C types capsular synthesizing enzymes and anticancer sequence on inducing the expression of apoptotic BCL-2, BAX, and Caspase-3 genes by Real-time RT-PCR method.

作者信息

Babakanrad Elmira, Mohammadian Taher, Esmaeili Davoud, Behzadi Payam

机构信息

Department of Microbiology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran.

Department of Microbiology and Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

出版信息

Heliyon. 2023 May 18;9(5):e16326. doi: 10.1016/j.heliyon.2023.e16326. eCollection 2023 May.

Abstract

BACKGROUND

Today, uterine cancer is one of the most important causes of death in the world and is one of the major problems in human health. There have been numerous reports of the effect of peptide and capsular products against cancer cell lines. : This study aimed to research recombinant peptide CPSA-CPSC-L-ACAN and investigate its apoptotic effect against the HeLa cell line by Real-Time-RT PCR.

DESIGN

In this study confirmation of the recombinant fusion peptide was performed by Western blotting. The effect of cytotoxicity of different concentrations of recombinant fusion peptide against the HeLa cell line was investigated by the MTT technique. The expression of apoptotic genes including BAX, BCL-2, and Caspase-3 in comparison with the GAPDH reference gene before and after exposure to recombinant fusion peptide was measured by Real-Time RT-PCR.

RESULTS

Recombinant fusion peptide at a concentration of 63 μg/ml destroyed 50% of the HeLa cell line in 24 h and cell treatment with this concentration increased gene expression of Caspase-3 genes by 16 times, bax by 6 times and decreased the expression of bcl-2 by 0.176 times.

CONCLUSIONS

The results showed that treatment of the HeLa cell line with recombinant fusion peptide induced an apoptotic effect. The recombinant fusion peptide could probably help the medical community as a prophylactic or therapeutic treatment for cervical cancer.

摘要

背景

如今,子宫癌是全球最重要的死亡原因之一,也是人类健康的主要问题之一。已有众多关于肽和包膜产品对癌细胞系作用的报道。本研究旨在研究重组肽CPSA-CPSC-L-ACAN,并通过实时逆转录聚合酶链反应(Real-Time-RT PCR)研究其对HeLa细胞系的凋亡作用。

设计

在本研究中,通过蛋白质免疫印迹法(Western blotting)对重组融合肽进行确认。采用MTT技术研究不同浓度重组融合肽对HeLa细胞系的细胞毒性作用。通过实时逆转录聚合酶链反应(Real-Time RT-PCR)测量与甘油醛-3-磷酸脱氢酶(GAPDH)参考基因相比,暴露于重组融合肽前后凋亡基因BAX、BCL-2和半胱天冬酶-3(Caspase-3)的表达。

结果

浓度为63μg/ml的重组融合肽在24小时内破坏了50%的HeLa细胞系,用该浓度处理细胞使Caspase-3基因的表达增加了16倍,bax增加了6倍,bcl-2的表达降低了0.176倍。

结论

结果表明,用重组融合肽处理HeLa细胞系可诱导凋亡作用。重组融合肽可能有助于医学界作为宫颈癌的预防性或治疗性治疗手段。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce5b/10213348/52bcf6c3c49f/gr1.jpg

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