胡桃醌对人宫颈癌HeLa细胞侵袭与迁移的影响

赵行宇 侯建成 金连海 王玉琪 刘嵩 张巍*

doi:10.16098/j.issn.0529-1356.2017.02.007

PDF(565 KB)

解剖学报 ›› 2017, Vol. 48 ›› Issue (2) : 160-164. DOI: 10.16098/j.issn.0529-1356.2017.02.007

肿瘤生物学

胡桃醌对人宫颈癌HeLa细胞侵袭与迁移的影响

赵行宇侯建成金连海王玉琪刘嵩张巍*

作者信息 +

Effect and its mechanism of Juglone on invasion and metastasis ability of human cervical cell line HeLa

ZHAO Xing-yu HOU Jian-cheng JIN Lian-hai WANG Yu-qi LIU Song ZHANG Wei*

Author information +

文章历史 +

摘要

目的观察胡桃醌对HeLa细胞侵袭及迁移的抑制作用，并探讨其可能的作用机制。方法常规体外培养人HeLa细胞，加入10、20、50、100μmol/L浓度的胡桃醌，继续培养24 h。通过倒置显微镜观察细胞形态学变化，采用MTT法检测细胞增殖情况，细胞划痕实验检测胡桃醌对细胞迁移能力的影响，用Transwell小室法检测细胞侵袭能力的影响。Western blotting检测基质金属蛋白酶（MMP）-2、MMP-9蛋白的表达。结果细胞形态学观察显示，不同浓度胡桃醌均可导致细胞形态学发生改变；MTT检测表明，胡桃醌对HeLa细胞的生长有明显抑制作用，且抑制作用随药物浓度的增加而增强(P<0.05或P<0.01)；划痕实验结果显示，随胡桃醌药物浓度的增加，细胞的平面运动能力明显下降；Transwell侵袭小室实验结果显示，胡桃醌对HeLa细胞体外的侵袭力具有很强的抑制作用；Western blotting结果表明，胡桃醌给药能够抑制HeLa细胞MMP-2、MMP-9蛋白的表达。结论胡桃醌能够抑制HeLa细胞的增殖并抑制细胞侵袭，其机制可能与抑制MMP-2、MMP-9的表达有关。

Abstract

Objective To observe the effect of Juglone on invasion and metastasis of Hela cells and explore the possible mechanism. Methods HeLa cells were cultured and treated with 10,20,50,100μmol/L Juglone for 24 hours. The morphology changes of HeLa cells were observed with an inverted microscope. The viability of HeLa cells was detected by MTT assay. The cell scratch test was used to detect cell migration ability after treatment of Juglone. The ability of cell invasion was measured by Transwell chamber. The expression of matrix metalloprateinases (MMP)-2 and MMP-9 were detected by Western blotting. Results Compared with control group, the viability of HeLa cells decreased after treatment with different concentrations of Juglone for 24 hours, and the cell morphology was changed in a dose-dependent manner. Scratch test results showed that the level of cell movement ability decreased significantly with the increase of the concentration of Juglone. Transwell invasion assay showed that Juglone had a strong inhibitory effect on the invasiveness of HeLa cells in vitro. Western blotting results showed that Juglone inhibited the expression of MMP-2 and MMP-9 protein in HeLa cells. Conclusion Juglone can inhibit the invasion and metastasis in HeLa cells, and its possible mechanism may be related to down regulating the expression of MMP-2 and MMP-9.

导出引用

赵行宇侯建成金连海王玉琪刘嵩张巍. 胡桃醌对人宫颈癌HeLa细胞侵袭与迁移的影响[J]. 解剖学报. 2017, 48(2): 160-164 https://doi.org/10.16098/j.issn.0529-1356.2017.02.007

ZHAO Xing-yu HOU Jian-cheng JIN Lian-hai WANG Yu-qi LIU Song ZHANG Wei. Effect and its mechanism of Juglone on invasion and metastasis ability of human cervical cell line HeLa[J]. Acta Anatomica Sinica. 2017, 48(2): 160-164 https://doi.org/10.16098/j.issn.0529-1356.2017.02.007

参考文献

［1］Gonzaleal MA,Tin A. Molecular biology of cervical cancer［J］. Clin Transl Oncol，2007，9(6):347-354.

［2］Gao Ye, Zhang J, Wu J, et al. Detection and genotyping of human papillom aviru sDNA in cervical cancer tissues with fluorescence polarization［J］. Acta Biochim Biophys Sin，2003，35(11 ): 1029-1034.

［3］Sugie S, Okamoto K, Rahman KM, et al. Inhibitory effects of plumbagin and juglone on azoxymethane-induced intestinal carcinogenesis in rats［J］. Cancer Lett, 1998,127(1):177-183.

［4］Chao SH, Greenleaf AL, Price DH. Juglone, an inhibitor of the peptidyl-prolyl isomerase Pin1, also directly blocks transcription［J］. Nucleic Acids Res, 2001,29(3):767-773.

［5］Varga Z, Bene L, Pieri C, et al. The effect of Juglone on the membrane potential and whole-cell K+ currents of human lymphocytes［J］. Biochemical and Biophysical Research Communications, 1996, 218(3):828-832.

［6］Xu HL, Yu XF, Qu SC, et al. Anti-proliferative effect of Juglone from Juglans mandshurica Maxim on human leukemia cell HL-60 by inducing apoptosis through the mitochondria-dependent pathway［J］. Eur J Pharmacol, 2010, 645(1-3):14-22.

［7］Xie J, Bai J, Sheng X, et al. Proliferation inhibition of human cervical cancer HeLa cells by Casticin in vitro［J］. The ChineseGerman Journal of Clinical Oncology，2011，10(1): 47-50.

［8］Malaguti C, Ciminiello P, Fattorusso E, et al. Caspase activation and death induced by yessotoxin in HeLa cells ［J］. Toxicol In Vitro, 2002, 16(4):357-363.

［9］Zhang W, Liu A, Li Y, et al. Anticancer activity and mechanism of juglone on human cervical carcinoma HeLa cells［J］. Can J Physiol Pharmacol, 2012,90(11):1553-1558.

［10］Lordick F， Hlscher AH， Haustermans K, et al. Multimodal treatment of esophageal cancer［J］. Langenbecks Arch Surg，2013，398(2):177-187.

［11］Ray ME，Mehra R，Sandler HM，et al. E-cadherin protein expression predicts prostate cancer salvage radiotherapy outcomes［J］. J Urol，2006，176(4 Pt1):1409-1414.

［12］Li W，Saji S，Sato F，et al. Potential clinical applications of matrix metalloproteinase inhibitors and their future prospects［J］. Int J Biol Markers，2013，28(2):e117-130.

［13］Jang ChZh,Chen JF,He LSh,et al. The fuction of MMP-2 and MMP-9 in primary Human brain gliomas invasion［J］. Acta Anatomica Sinica, 2001,32(4):346-349. (in Chinese)

江常震, 陈锦峰, 何理盛, 等. MMP-2和MMP-9在人脑原发性胶质瘤侵袭中的作用［J］.解剖学报,2001,32(4):346-349.

［14］Wang XJ,Yang YJ,Tian HN,et al. Effects of annexin A5 low expression on migration and invasion of HeLa cells［J］. Acta Anatomica Sinica, 2016，47(4):487-492. . (in Chinese)王小杰, 杨艳杰,田焕娜,等.膜联蛋白 A5 低表达对人宫颈癌 HeLa 细胞迁移和侵袭的影响［J］. 解剖学报，2016，47(4):487-492.

［15］Radenkovic S，Konjevic G，Jurisic V，et al. Values of MMP-2 and MMP-9 in tumor tissue of basal-like breast cancer patients［J］. Cell Biochem Biophys，2014，68(1): 143-152.

［16］Shao W，Wang W，Xiong XG，et al. Prognostic impact of MMP-2 and MMP-9 expression in pathologic stage IA non-small cell lung cancer［J］. J Surg Oncol，2011，104(7): 841-846.