低强度超声对肝癌细胞MHCC97H迁移与侵袭能力影响的体外研究

doi:10.3969/j.issn.1007-3969.2015.12.002

中国癌症杂志 ›› 2015, Vol. 25 ›› Issue (12): 926-932.doi: 10.3969/j.issn.1007-3969.2015.12.002

低强度超声对肝癌细胞MHCC97H迁移与侵袭能力影响的体外研究

杨名珍，刘邦忠，石明芳，李　蕴，刘光华，王　平

复旦大学附属中山医院康复医学科，上海 200032

出版日期:2015-12-30 发布日期:2016-02-03
通信作者: 刘邦忠　E-mail:liu.bangzhong@zs-hospital.sh.cn

An in vitro study of the effect of low intensity ultrasound on the migration and invasion of MHCC97H

YANG Mingzhen, LIU Bangzhong, SHI Mingfang, LI Yun, LIU Guanghua, WANG Ping

Department of Rehabilitation, Zhongshan Hospital, Fudan University, Shanghai 200032, China

Published:2015-12-30 Online:2016-02-03
Contact: LIU Bangzhong E-mail: liu.bangzhong@zs-hospital.sh.cn

摘要/Abstract

摘要： 背景与目的：低强度超声(low intensity ultrasound, LIUS)能杀伤肿瘤细胞，促进肿瘤细胞的凋亡，但对肿瘤细胞迁移侵袭作用的影响仍不清楚。该研究旨在探讨LIUS对肝癌细胞MHCC97H迁移侵袭的影响及可能的机制。方法：实验根据超声照射强度的不同分为对照组(0 W/cm²)、0.5 W/cm²组、1.0 W/cm²组和1.5 W/cm²组；LIUS处理后划痕实验和Transwell小室体外迁移和侵袭实验检测肿瘤细胞的迁移和侵袭能力；显微镜及F-actin细胞骨架荧光染色观察处理后细胞骨架的变化；实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction，RTFQ-PCR)及蛋白质［印］迹法(Western blot)检测各组MMP-2、MMP-9的表达变化。结果：LIUS(小于等于1.5 W/cm²)能促进肝癌细胞MHCC97H的迁移侵袭，划痕实验及Transwell迁移和侵袭实验都表明LIUS处理后肿瘤细胞迁移侵袭数量增加；光学显微镜及荧光显微镜发现LIUS处理后肝癌细胞的形态发生变化；RTFQ-PCR及Western blot的结果表明超声处理后MMP-2 mRNA 及蛋白表达水平上升，MMP-9的mRNA 表达水平上升。结论：LIUS可能通过改变肿瘤细胞的骨架及增加MMP-2的表达促进肝癌细胞MHCC97H的迁移及侵袭能力。

关键词: 低强度超声, 肝癌细胞, 转移\基质金属蛋白酶

Abstract: Background and purpose: Low intensity ultrasound (LIUS) can kill cancer cells and promote their apoptosis. However, it is still unknown how it affects the migration and invasion of tumor cells. This study aimed to explore the effect of LIUS on human hepatocellular line MHCC97H in migration and metastasis and the possible mechanism in vitro. Methods: According to the intensity of ultrasonic irradiation, 4 experimental groups were established: control group (0 W/cm²), 0.5 W/cm², 1.0 W/cm² and 1.5 W/cm²group. The migration and invasion ability of hepatocellular cells was detected by scratch assay and Transwell migration and invasion assay after the irradiation of LIUS. The changes of cytoskeleton after irradiation were observed by microscope and F-actin green fluorescence staining. The expressions of MMP-2 and MMP-9 were examined by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Results: Low intensity ultrasound (≤1.5 W/cm²) promoted the migration and invasion of hepatocellular line MHCC97H. Scratch assay and Transwell assay showed much more cells under irradiation migrated through membrane than untreated. It was found that morphology of liver cancer cells changed after LIUS irradiation using optical microscope and fluorescence microscope. The results of RTFQ-PCR and Western blot showed upregulation of MMP-2 expression by LIUS in MHCC97H and high expression of MMP-9 mRNA. Conclusion: Low intensity ultrasound may promote the migration and invasion of MHCC97H through changing cytoskeleton and upregulating protein expression of MMP-2.

Key words: Low intensity ultrasound, Liver cancer cell, Migration, Matrix metalloproteinase

杨名珍,刘邦忠,石明芳,等. 低强度超声对肝癌细胞MHCC97H迁移与侵袭能力影响的体外研究[J]. 中国癌症杂志, 2015, 25(12): 926-932.

YANG Mingzhen, LIU Bangzhong, SHI Mingfang, et al. An in vitro study of the effect of low intensity ultrasound on the migration and invasion of MHCC97H[J]. China Oncology, 2015, 25(12): 926-932.

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低强度超声对肝癌细胞MHCC97H迁移与侵袭能力影响的体外研究

An in vitro study of the effect of low intensity ultrasound on the migration and invasion of MHCC97H

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