Effects of miR-125b-5p on the proliferation and apoptosis of human hemangioma endothelial cells HemES and its mechanism

doi:10.12047/j.cjap.6068.2021.021

Abstract

Abstract: Objective: This article mainly studies the effects of miR-125b-5p on the proliferation and apoptosis of human hemangioma endothelial cells HemECs. Methods: RT-qPCR was used to detect the expressions of miR-125b-5p and MCL-1 mRNA in HemECs and collateral cells of human hemangioma endothelial cells. HemECs were selected and divided into control group, miR-NC group, miR-125b-5p mimic group, miR-125b-5p inhibitor group, pc-MCL-1 group, miR-125b-5p+ pc-MCL-1 group, 9 multiple holes in each group. . HemECs were transfected with 100 nmol · L^-1 of miR-NC, miR-125b-5p mimic, miR-125b-5p inhibitor, pc-MCL-1 plasmids separately or in combination. MTT method was used to detect the proliferation of HemECs. The apoptosis of HemECs was detected by flow cytometry. Dual luciferase report was used to to detect targeting relationship. The relative expression levels of Ki67, PCNA, cleaved caspase-3, Bax, Bcl-2, p-p70s6k/ p70s6k, p-AKT/AKT, and p-mTOR/mTOR proteins were detected by Western blot. Results: By comparing the expression levels of miR-125b-5p in hemangioma tissues and cells, HemECs cell lines with obvious down-regulation effects were selected for follow-up experiments. Compared with the control group, the proliferation of HemECs and the expressions of Ki67 and PCNA in the miR-125b-5p mimic group were decreased significantly (P＜0.01). The apoptotic rate of HemECs and the expression levels of cleaved Caspase-3 and Bax were increased significantly, while the expression of Bcl-2 was decreased significantly (P＜0.01). The expression levels of p-AKT/AKT, p-mTOR/mTOR and p-p70S6K/p70S6K were down-regulated significantly (P ＜0.01); the proliferation of HemECs and the expressions of Ki67 and PCNA in the miR-125b-5p inhibitor group were increased significantly (P ＜0.01); the apoptosis rate and the expressions of cleaved Caspase-3 and Bax were decreased significantly, and the expression of Bcl-2 was increased (P＜0.05, P＜0.01). miR-125b-5p targeted down-regulation of MCL-1. Compared with miR-125b-5p mimic group, the proliferation of HemECs and the expressions of Ki67 and PCNA in miR-125b-5p+ pc-MCL-1 group were increased significantly (P＜0.01), the apoptosis rate of HemECs and the expressions of cleaved Caspase-3 and Bax were decreased significantly, while the expression of Bcl-2 was increased (P＜0.01). The expressions of p-AKT/AKT, p-mTOR/mTOR, and p-p70S6K/p70S6K was also increased significantly (P＜0.01). Conclusion: miR-125b-5p inhibits the proliferation of human hemangioma endothelial cells and induces apoptosis. The mechanism may be related to the targeted down-regulation of MCL-1 expression and inhibition of AKT/mTOR pathway activation.

Key words: miR-125b-5p, MCL-1, hemangioma, proliferation, apoptosis

CLC Number:

R732.2

WANG Tian, ZHANG Fu-lin, ZHAO Yue, GUO Dong-dong, YANG Rui. Effects of miR-125b-5p on the proliferation and apoptosis of human hemangioma endothelial cells HemES and its mechanism[J]. CJAP, 2021, 37(3): 247-253.

References 19

[1]	Auger N, Fraser WD, Arbour L, et al. Pre-eclampsia and risk of infantile haemangioma [J]. Br J Dermatol, 2017, 176(2): 371-377.
[2]	陈玲玲, 冯珊珊, 范祖森, 等. 非编码RNA研究进展[J]. 中国科学: 生命科学, 2019, 49(12): 1573-1605.
[3]	Sun C, Wang XM, Cao HR, et al. Regulation of mir-338-3p targeting tcf4 on the proliferation, apoptosis and metastasis of human renal carcinoma cells [J]. J Med Mol Bio, 2019, 16 (1): 62-68.
[4]	Huang C, Huang J, Ma P, et al. Microrna-143 acts as a suppressor of hemangioma growth by targeting bcl-2 [J]. Gene, 2017, 628(2): 211-217.
[5]	Huang J, Jiang D, Zhao S, et al. Propranolol suppresses infantile hemangioma cell proliferation and promotes apoptosis by upregulating mir-125b expression [J]. Anticancer Drugs, 2019, 30(5): 501-507.
[6]	孙凤娥, 方浩. Mcl-1蛋白的结构、功能及其抑制剂的研究进展[J]. 中国医药工业杂志, 2013, 44(3): 296-302.
	Liu C, Zhao Z, Ji Z, et al. Mir-187-3p enhances propranolol sensitivity of hemangioma stem cells [J]. Cell Struct Funct, 2019, 44(1): 41-50.
[7]	Zheng Z, Qu JQ, Yi HM, et al. Mir-125b regulates proliferation and apoptosis of nasopharyngeal carcinoma by targeting a20/nf-κb signaling pathway [J]. Cell Death Dis, 2017, 8(6): e2855.
[8]	Wu S, Liu F, Xie L, et al. Mir-125b suppresses proliferation and invasion by targeting mcl1 in gastric cancer [J]. Biomed Res Int, 2015, 15(2): 365273.
[9]	Wang Y, Tang P, Chen Y, et al. Overexpression of microrna-125b inhibits human acute myeloid leukemia cells invasion, proliferation and promotes cells apoptosis by targeting nf-κb signaling pathway [J]. Biochem Biophys Res Commun, 2017, 488(1): 60- 66.
[10]	Liu S, Chen Q, Wang Y. Mir-125b-5p suppresses the bladder cancer progression via targeting hk2 and suppressing pi3k/akt pathway [J]. Hum cell, 2019, 10(11): 285- 294.
[11]	Li Y, Wang Y, Fan H, et al. Mir-125b-5p inhibits breast cancer cell proliferation, migration and invasion by targeting kiaa1522 [J]. Biochem Biophys Res Commun, 2018, 504(1): 277-282.
[12]	Zhang D, Yi Z, Fu Y. Downregulation of mir-20b-5p facilitates mycobacterium tuberculosis survival in raw 264.7 macrophages via attenuating the cell apoptosis by mcl-1 upregulation [J]. J Cell Biochem, 2019, 120(4): 5889-5896.
[13]	Feng WT, Yao R, Xu LJ, et al. Effect of mir-363 on the proliferation, invasion and apoptosis of laryngeal cancer by targeting mcl-1 [J]. Eur Rev Med Pharmacol Sci, 2018, 22(14): 4564-4572.
[14]	Zhang T, Zou P, Wang T, et al. Down-regulation of mir-320 associated with cancer progression and cell apoptosis via targeting mcl-1 in cervical cancer [J]. Tumour Biol, 2016, 37(7): 8931-8940.
[15]	Lou S, Wang Y, Yu Z, et al. Curcumin induces apoptosis and inhibits proliferation in infantile hemangioma endothelial cells via downregulation of mcl-1 and hif-1α [J]. Medicine, 2018, 97(7): e9562.
[16]	Xu X, Jiang J, Yao L, et al. Silencing the folr2 gene inhibits cell proliferation and increases apoptosis in the nci-h1650 non-small cell lung cancer cell line via inhibition of akt/mammalian target of rapamycin (mtor)/ribosomal protein s6 kinase 1 (s6k1) signaling [J]. Med Sci Monit, 2018, 24(11): 8064-8073.
[17]	Liang QP, Xu TQ, Liu BL, et al. Sasanquasaponin ιιι from schima crenata korth induces autophagy through akt/mtor/p70s6k pathway and promotes apoptosis in human melanoma a375 cells [J]. Phytomedicine, 2019, 58(2): 152769.
[18]	Zhu J, Yao J, Huang R, et al. Ghrelin promotes human non-small cell lung cancer a549 cell proliferation through pi3k/akt/mtor/p70s6k and erk signaling pathways [J]. Biochem Biophys Res Commun, 2018, 498(3): 616-620.
[19]	Li D, Li P, Guo Z, et al. Downregulation of mir-382 by propranolol inhibits the progression of infantile hemangioma via the pten-mediated akt/mtor pathway [J]. Int J Mol Med, 2017, 39(3): 757-763.