南蛇藤醇抑制宫颈癌细胞生物学行为的机制研究

贾莹, 赵伟, 张欣媛, 吴瑞, 肖兵兵

doi:10.13418/j.issn.1001-165x.2020.04.011

中国临床解剖学杂志 >

2020 , Vol. 38 >Issue 4: 421 - 427

DOI: https://doi.org/10.13418/j.issn.1001-165x.2020.04.011

实验研究

南蛇藤醇抑制宫颈癌细胞生物学行为的机制研究

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1.大庆龙南医院（齐齐哈尔医学院第五附属医院），黑龙江大庆 163000； 2.大庆油田总医院
临床药学科，黑龙江大庆 163316； 3.大庆市人民医院妇产科，黑龙江大庆 163000

贾莹（1986-），女，山东人，硕士，主要从事药学研究工作，E-mail：jy86210@126.com

收稿日期: 2019-06-14

网络出版日期: 2020-07-29

基金资助

黑龙江省卫生和计划生育委员会（【2018】129）

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The mechanism of Celastrol in inhibiting cell biological behavior of cervical cancer cells

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1. Daqing Longnan Hospital (the Fifth Hospital of Qiqihar Medical University), Daqing 163000, Heilongjiang Province, China; 2. Department of Clinical Pharmacy, General Hospital of Daqing Oilfield 163316, Heilongjiang Province, China; 3. Department of Obstetrics and Gynecology, Daqing People's Hospital 163000, Heilongjiang Province, China

Received date: 2019-06-14

Online published: 2020-07-29

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摘要

目的　探讨南蛇藤醇抑制宫颈癌细胞增殖和侵袭的作用机制。方法　将正常培养HeLa细胞分为对照组、南蛇藤醇低、中、高剂量组（终浓度分别为4 μmol/L、8 μmol/L和12 μmol/L）、阳性对照组（顺铂，终浓度为10 μmol/L）、LY294002组（LY294002，终浓度为20 μmol/L）和（南蛇藤醇+LY294002）组（南蛇藤醇和LY294002，终浓度分别为12 μmol/L和20 μmol/L）。MTT法检测各组细胞存活情况；流式细胞仪检测细胞凋亡情况；Transwell检测细胞的侵袭；蛋白质印迹法检测PI3K/AKT/NF-κB信号通路的表达；采用皮下注射HeLa细胞建立裸鼠模型，观察南蛇藤醇（120 mg/kg）对裸鼠移植瘤体积和重量的影响。结果　与对照组相比，南蛇藤醇组、阳性对照组和LY294002组HeLa细胞的相对增殖率、侵袭细胞数均明显下降（P<0.001），细胞凋亡率明显升高（P<0.001），p-PI3K、p-AKT、NF-κB p65蛋白表达明显下调（P<0.001），且随着南蛇藤醇浓度升高，其作用增强；与LY294002组相比，（南蛇藤醇+LY294002）组HeLa细胞的相对增殖率和侵袭细胞数明显下降（q=10.182，q=10.217，P均<0.001），细胞凋亡率明显升高（q=23.636，P<0.001）；与对照组相比，南蛇藤醇组裸鼠体重、移植瘤的体积和重量明显下降（q=4.100，P<0.020；q=13.501，P<0.001；q=5.078，P=0.005）。结论　南蛇藤醇可能通过抑制HeLa细胞的PI3K/Akt/NF-κB信号通路，抑制其恶性生物学行为。

关键词： 南蛇藤醇; 　宫颈癌; 　PI3K/AKT/NF-κB信号通路

本文引用格式

贾莹, 赵伟, 张欣媛, 吴瑞, 肖兵兵 . 南蛇藤醇抑制宫颈癌细胞生物学行为的机制研究[J]. 中国临床解剖学杂志, 2020 , 38(4) : 421 -427 . DOI: 10.13418/j.issn.1001-165x.2020.04.011

Abstract

Objective　To explore the mechanism of Celastrol in inhibiting proliferation and invasion ability of cervical cancer cells.　Methods　Normal cultured HeLa cells were divided into the following groups: a control group, a low-dose Celastrol group, a medium-dose Celastrol group and a high-doses Celastrol group (final concentrations of Celastrol were 4 μmol/L, 8 μmol/L and 12 μmol/L, respectively), a positive control group (cisplatin, final concentration of 10 μmol/L), a LY294002 group (LY294002, final concentration of 20 μmol/L) and a Celastrol with LY294002 group ( final concentrations of Celastrol and LY294002 were 12 μmol/L and 20 μmol/L, respectively). The cell survival conditions of each group were detected by MTT assay. The apoptosis of each group were detected by flow cytometry. The cell invasion of each group were detected by Transwell. The expression of PI3K/AKT/NF-κB signaling pathway were detected by Western Blot. HeLa cells were subcutaneously injected to establish nude mice models to observe the effects of Celastrol (120 mg/kg) on volume and weight of transplanted tumors in nude mice.　Results　Compared with the control group, relative proliferation rate of HeLa cells and number of invasive cells in Celastrol group, positive control group and LY294002 group significantly decreased (P<0.001), while apoptosis rates significantly increased (P<0.001), and expression of p-PI3K, p-AKT and NF-κB p65 protein significantly down-regulated (P<0.001). Moreover, with increasing of Celastrol concentration, its effects increased. Compared with the LY294002 group, relative proliferation rate of HeLa cells and number of invasive cells in the Celastrol with LY294002 group significantly decreased (q=10.182, q=10.217; P<0.001), while apoptosis rate significantly increased (q=23.636, P<0.001). Compared with the control group, weight of nude mice, volume and weight of transplanted tumors of nude mice significantly decreased in the Celastrol groups (q=4.100, P<0.020; q=13.501, P<0.001; q=5.078, P=0.005).　Conclusions　Celastrol may inhibit its malignant biological behaviors through inhibiting the PI3K/Akt/NF-κB signaling pathway of HeLa cells

Key words： Celastrol; 　Cervical cancer; 　PI3K/AKT/NF-κB signaling pathway

参考文献

[1] Kumakech E, Andersson S, Wabinga H, et al. Cervical cancer risk perceptions, sexual risk behaviors and sexually transmitted infections among Bivalent Human Papillomavirus vaccinated and non-vaccinated young women in Uganda-5 year follow up study[J]. Bmc Womens Health, 2017, 17(1): 40-51.
[2] Wolf B, Ganzer R, Stolzenburg JU, et al. Extended mesometrial resection (EMMR): surgical approach to the treatment of locally advanced cervical cancer based on the theory of ontogenetic cancer fields[J]. Gynecol Oncol, 2017, 146(2): 292-298.
[3] 王艳萍, 徐静, 张丽钰, 等. 中药清毒栓对宫颈癌SiHa细胞抑制作用的影响[J]. 中国老年学杂志, 2016, 36(8): 1822-1825.
[4] 陈光福, 郭远清, 伍玉甜. 南蛇藤素可抑制肿瘤坏死因子α诱导RAW264.7细胞的炎症及增殖[J]. 中国组织工程研究, 2016, 20(37): 5552-5559.
[5] 钱亚云, 李文原, 赵雪煜, 等. 南蛇藤提取物靶向mTOR抑制人肝癌HepG2细胞侵袭与转移能力的研究[J]. 中草药, 2018, 49(20): 4831-4837.
[6] Bedell S, Manders D, Kehoe S, et al. The opinions and practices of providers toward the sexual issues of cervical cancer patients undergoing treatment[J]. Gynecol Oncol, 2017, 144(3): 586-591.
[7] Orang'O EO, Itsura P, Tonui P, et al. Use of palliative cisplatinum for advanced cervical cancer in a resource-poor setting: a case series from kenya[J]. J Glob Oncol, 2016, 3(5): 539-544.
[8] 王海波, 顾昊, 赵雪煜, 等. 南蛇藤提取物通过调控基质金属蛋白酶组及其抑制因子抑制人胃癌SGC-7901细胞侵袭转移的研究[J]. 中草药, 2016, 47(8): 1345-1350.
[9] Zhou YQ, Li WC, Wang ML, et al. Competitive profiling of celastrol targets in human cervical cancer HeLa cells via quantitative chemical proteomics[J]. Mol Biosyst, 2016, 13(1): 83-91.

[10]Ciepliński K, Jóźwik M, Semczuk-Sikora A, et al. Expression of p53 and selected proliferative markers (Ki-67, MCM3, PCNA, and topoisomerase IIα) in borderline ovarian tumors: correlation with clinicopathological features[J]. Histol Histopathol,2018, 33(2):171-179.

[11]Wang SJ, Dong LQ, Liu H, et al. Effect of celastrol on anti-proliferation and energy metabolism in SGC-7901 cells and ECV304 cells[J]. Chin Tradit Herbal Drugs, 2016, 47(21): 3854-3860.
[12] Wang J, Yang DJ, Luo QY, et al. APG-1252-12A induces mitochondria-dependent apoptosis through inhibiting the antiapoptotic proteins Bcl-2/Bcl-xl in HL-60 cells[J]. Int J Oncol, 2017, 51(2): 563-572.
[13]Li PP, He W, Yuan PF, et al. Celastrol induces mitochondria-mediated apoptosis in hepatocellular carcinoma Bel-7402 cells[J]. Am J Chin Med, 2015, 43(1):137-148.
[14] Zhao ZT, Fan XY, Jiang LF, et al. miR-503-3p promotes epithelial-mesenchymal transition in breast cancer by directly targeting SMAD2 and E-cadherin[J]. J Genet Genomics, 2017, 44(2): 75-84.
[15]Mo DP, Yang DH, Xiao XL, et al. MiRNA-145 suppresses lung adenocarcinoma cell invasion and migration by targeting N-cadherin[J]. Biotechnol Lett, 2017, 39(5): 701-710.
[16]Labernadie A, Kato T, Brugués A, et al. A mechanically active heterotypic E-cadherin/N-cadherin adhesion enables fibroblasts to drive cancer cell invasion[J]. Nat Cell Biol, 2017, 19(3): 224-237.
[17] Wu JH, Ding MC, Mao NF, et al. Celastrol inhibits chondrosarcoma proliferation, migration and invasion through suppression CIP2A/c-MYC signaling pathway[J]. J Pharmacol Sci, 2017, 134(1): 22-28.
[18]Prue G, Lawler M, Baker P, et al. Human papillomavirus (HPV): making the case for ‘Immunisation for All’[J]. Oral Dis, 2017, 23(6): 726-730.
[19] Chen HH, Chen SP, Zheng QL, et al. Genistein promotes proliferation of Human cervical cancer cells through estrogen receptor-mediated PI3K/Akt-NF-κB pathway[J]. J Cancer, 2018, 9(2): 288-295.
[20] Yang L, Liu YQ, Wang M, et al. Celastrus orbiculatus extract triggers apoptosis and autophagy via PI3K/Akt/mTOR inhibition in human colorectal cancer cells[J]. Oncol Lett, 2016, 12(5): 3771-3778.
[21]Fu X, Wen HQ, Jing L, et al. MicroRNA-155-5p promotes hepatocellular carcinoma progression by suppressing PTEN through the PI3K/Akt pathway[J]. Cancer Sci, 2017, 108(4): 620-631.
[22] Grinberg-Bleyer Y, Oh H, Desrichard A, et al. NF-κB c-Rel is crucial for the regulatory T cell immune checkpoint in cancer[J]. Cell, 2017, 170(6): 1096-1108.
[23] Yu XL, Wang Q, Zhou X, et al. Celastrol negatively regulates cell invasion and migration ability of human osteosarcoma via downregulation of the PI3K/Akt/NF-κB signaling pathway in vitro[J]. Oncol Lett, 2016, 12(5): 3423-3423.
[24] Tang BF, Xu D, Zhao YL, et al. Celastrol inhibits colorectal cancer cell proliferation and migration through suppression of MMP3 and MMP7 by the PI3K/AKT signaling pathway[J]. Anticancer Drugs, 2018, 29(6): 530-538.

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