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微小 RNA miR-30a 通过自噬抑制卵巢癌细胞对顺铂的耐药性。

MicroRNA miR-30a inhibits cisplatin resistance in ovarian cancer cells through autophagy.

机构信息

Department of Oncology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China.

出版信息

Bioengineered. 2021 Dec;12(2):10713-10722. doi: 10.1080/21655979.2021.2001989.

DOI:10.1080/21655979.2021.2001989
PMID:34747309
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8810079/
Abstract

We study whether microRNA miR-30a inhibits the autophagy through transforming growth factor (TGF)-β/Smad4 to generate cisplatin (DDP) resistance in ovarian cancer cells. The expression of miR-30a, Smad4, and TGF-β was detected in the serum of ovarian cancer patients and DDP-resistant cell lines (A2780) by quantitative real-time polymerase chain reaction (qRT-PCR). Computational search and western blotting were used to demonstrate the downstream target of miR-30a in ovarian cancer cells. Cell viability was measured with CCK8 assay. Apoptosis and autophagy of ovarian cancer cells were analyzed by flow cytometry and transmission electron microscopy, and the expressions of Beclin1 and LC3II protein were detected by western blotting. Expression of miR-30a was significantly decreased, while expressions of TGF-β and Smad4 mRNA were increased in serum of ovarian cancer patients after DDP chemotherapy as well as in DDP-resistant cells. Activation of autophagy contributed to DDP-resistance cells. Moreover, Bioinformatics software predicted Smad4 to be a target of miR-30a. Overexpression of miR-30a decreased the expression of Smad4 and TGF-β. Additionally, miR-30a-overexpressing inhibited DDP-induce autophagy and promoted DDP-resistant cell apoptosis. In conclusion, miR-30a mediates DDP resistance in ovarian cancer by inhibiting autophagy via the TGF-β/Smad4 pathway.

摘要

我们研究了 microRNA miR-30a 是否通过转化生长因子 (TGF)-β/Smad4 抑制自噬来产生卵巢癌细胞对顺铂 (DDP) 的耐药性。通过定量实时聚合酶链反应 (qRT-PCR) 检测卵巢癌患者血清和 DDP 耐药细胞系 (A2780) 中 miR-30a、Smad4 和 TGF-β 的表达。计算搜索和 Western blot 用于证明 miR-30a 在卵巢癌细胞中的下游靶标。用 CCK8 测定法测量细胞活力。通过流式细胞术和透射电子显微镜分析卵巢癌细胞的凋亡和自噬，并通过 Western blot 检测 Beclin1 和 LC3II 蛋白的表达。在 DDP 化疗后以及 DDP 耐药细胞中，卵巢癌患者血清中 miR-30a 的表达明显降低，而 TGF-β 和 Smad4 mRNA 的表达增加。自噬的激活有助于 DDP 耐药细胞。此外，生物信息学软件预测 Smad4 是 miR-30a 的靶标。miR-30a 的过表达降低了 Smad4 和 TGF-β 的表达。此外，miR-30a 过表达抑制 DDP 诱导的自噬并促进 DDP 耐药细胞凋亡。总之，miR-30a 通过 TGF-β/Smad4 通路抑制自噬介导卵巢癌对 DDP 的耐药性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc2f/8810079/111cb97868c0/KBIE_A_2001989_F0006_OC.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc2f/8810079/d0674ee706d5/KBIE_A_2001989_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc2f/8810079/c7c5a71e64d2/KBIE_A_2001989_F0002_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc2f/8810079/890ab5003816/KBIE_A_2001989_F0003_B.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc2f/8810079/111cb97868c0/KBIE_A_2001989_F0006_OC.jpg

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