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重组可溶性突变型白细胞介素-6受体对卵巢癌细胞系细胞毒性作用的评估

Evaluation of the Cytotoxic Effects of a Recombinant form of the Soluble Mutant IL-6 Receptor on an Ovarian Cancer Cell Line.

作者信息

Khatir Kosar, Aghaei Mahmoud, Ghorbanhosseini Seyedeh Sara, Shafiee Fatemeh

机构信息

Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Iran J Biotechnol. 2025 Jan 1;23(1). doi: 10.30498/ijb.2025.467140.3953. eCollection 2025 Jan.

DOI:10.30498/ijb.2025.467140.3953
PMID:40463947
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12128949/
Abstract

BACKGROUND

Interleukin-6 (IL-6) plays an important role in cancer metastasis. Therefore, it seems that inhibition of IL-6 may act as an anticancer agent.

OBJECTIVES

This study examined the anti-proliferative and IL-6 signal transduction inhibitory effects of a mutant IL-6 receptor (mIL-6R) on an ovarian cancer cell line.

MATERIALS AND METHODS

The intein1-mIL-6R was expressed in BL21 at various inducer concentrations and temperatures. The expressed protein was purified using the IMPACT system, and the best-examined conditions (i.e., temperature, time, cleavage buffer pH) for intein1auto-cleavage were achieved. The anti-proliferative effects of mIL-6R on OVCAR-3 cancer cells were investigated using MTT assay and RT-PCR to determine its effects on suppressing the JAK-STAT pathway genes.

RESULTS

The highest solubility of mIL-6R was obtained at 20 °C, 0.5 mM IPTG. The highest level of intein1 cleavage occurred at 25 °C, 24 hours of incubation, and pH = 4 of cleavage buffer. Also, mIL-6R diminished the survival of OVCAR-3 cells compared to PBS (p-value = 0.024), with 48 hours IC50 of 1.117 μg/mL. Also, mIL-6R significantly reduced the expression of the JAK, STAT, and VEGF genes.

CONCLUSION

The recombinant mIL-6R showed a strong concentration-dependent anti-proliferative effect on the OVCAR-3 cell line. However, it needs further evaluation for its potential activity against disorders associated with increased IL-6.

摘要

背景

白细胞介素-6（IL-6）在癌症转移中起重要作用。因此，抑制IL-6似乎可作为一种抗癌剂。

目的

本研究检测了突变型IL-6受体（mIL-6R）对卵巢癌细胞系的抗增殖和IL-6信号转导抑制作用。

材料与方法

在不同诱导剂浓度和温度下，在BL21中表达内含肽1-mIL-6R。使用IMPACT系统纯化表达的蛋白质，并确定内含肽1自切割的最佳检测条件（即温度、时间、切割缓冲液pH值）。使用MTT法研究mIL-6R对OVCAR-3癌细胞的抗增殖作用，并通过RT-PCR确定其对抑制JAK-STAT途径基因的影响。

结果

在20℃、0.5 mM IPTG条件下，mIL-6R的溶解度最高。内含肽1的最高切割水平发生在25℃、孵育24小时、切割缓冲液pH = 4时。此外，与PBS相比，mIL-6R降低了OVCAR-3细胞的存活率（p值 = 0.024），48小时IC50为1.117μg/mL。此外，mIL-6R显著降低了JAK、STAT和VEGF基因的表达。

结论

重组mIL-6R对OVCAR-3细胞系显示出强烈的浓度依赖性抗增殖作用。然而，其对与IL-6升高相关疾病的潜在活性需要进一步评估。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/c4b1f962c9c3/IJB-23-e3953-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/be40ad35ebf4/IJB-23-e3953-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/f0de88614c83/IJB-23-e3953-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/6deb02d33b6f/IJB-23-e3953-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/c4b1f962c9c3/IJB-23-e3953-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/be40ad35ebf4/IJB-23-e3953-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/f0de88614c83/IJB-23-e3953-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/6deb02d33b6f/IJB-23-e3953-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/089f/12128949/c4b1f962c9c3/IJB-23-e3953-g004.jpg

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本文引用的文献

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