特异性下调LKB1的表达对肝癌细胞侵袭迁移能力的影响

沙亮; 何剑波; 李科志; 陈闯; 黄山; 赵荫农; 连芳; 邬国斌

doi:10.3969/j.issn.1001-5256.2017.05.016

特异性下调LKB1的表达对肝癌细胞侵袭迁移能力的影响

DOI: 10.3969/j.issn.1001-5256.2017.05.016

1. 广西医科大学附属肿瘤医院肝胆外科
2. 广西医科大学研究生院
3. 广西医科大学基础医学院

基金项目:

国家自然科学基金资助项目(81460426);国家自然科学基金资助项目(81360315)；

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中图分类号: R735.7
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出版历程
- 出版日期: 2017-05-20

Influence of specific down-regulation of LKB1 expression on invasion and migration of hepatoma cells

Department of Hepatobiliary Surgery, The Affiliated Tumor Hospital of Guangxi Medical University

Research funding:

摘要

摘要:
目的研究特异性下调LKB1的表达对肝癌细胞系SK-hep-1迁移侵袭能力的影响。方法 Real-time PCR和Western Blot检测人肝癌细胞株和人正常肝细胞HL7702中LKB1表达情况。设计并合成LKB1特异性双链小干扰RNA（LKB1-siRNA）,转染到高表达LKB1的人肝癌细胞株SK-hep-1中靶向沉默LKB1基因表达,并设置阴性对照组（NC组）。Real-time PCR和Western Blot检测LKB1基因的mRNA水平与蛋白表达水平的变化。利用CCK-8法检测肝癌细胞增殖能力。通过体外Transwell小室基质侵袭和迁移实验,观察LKB1表达沉默后对人肝癌细胞株SK-hep-1侵袭迁移能力的影响。计量资料两组间比较采用t检验,多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与正常人肝细胞HL7702相比,LKB1基因和蛋白在人肝癌细胞SK-hep-1中高表达（P值均<0.05）。与NC组相比,siLKB1-1处理组SK-hep-1细胞中的LKB1基因和蛋白表达均明显降低（P值均<0.05）。与NC组相比,siLKB1-1转...
- 癌,肝细胞 /
- LKB1 /
- 细胞增殖 /
- 肿瘤侵润 /
- 细胞运动
Abstract:
Objective To investigate the influence of specific down-regulation of LKB1 expression on invasion and migration of the hepatoma cell line SK-hep-1.Methods Quantitative real-time PCR (RT-PCR) and Western blot were used to measure the expression of LKB1 in the human hepatoma cell line SK-hep-1 and normal hepatocytes (HL7702) .LKB1 specific double-strand small interfering RNA was designed, synthesized, and then transfected into the human hepatoma cell line SK-hep-1 with high expression of LKB1 to silence the expression of LKB1 gene.A negative control (NC) group was established.RT-PCR and Western blot were used to measure the changes in the mRNA and protein expression of LKB1.Cell Counting Kit-8 was used to measure the proliferative capacity of hepatoma cells.In vitro Transwell chamber invasion and migration assays were used to observe the influence of LKB1 gene silencing on invasion and migration of the human hepatoma cell line SK-hep-1.The t-test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between any two groups.Results Compared with normal human hepatocytes (HL7702) , SK-hep-1 cells had significantly higher mRNA and protein expression of LKB1 (both P<0.05) .Compared with the NC group, the siLKB1-1 treatment group had significant reductions in the mRNA and protein expression of LKB1 in SK-hep-1 cells (both P<0.05) .Compared with the NC group, the siLKB1-1 treatment group had a significantly higher cell proliferation rate and significant increases in invasion and migration (1.393 ±0.022 vs 1.1284±0.032, t=15.313, P<0.001;147±19 vs 83±21, t=14.879, P<0.001;113±13 vs 75±17, t=8.791, P<0.001) .Conclusion LKB1 may be involved in inhibiting the proliferation, invasion, and migration of hepatoma cells and can affect the prognosis of patients with liver cancer.
- carcinoma, hepatocellular /
- liver kinase b1 /
- cell proliferation /
- neoplasm invasiveness /
- cell movement

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参考文献(19)

[1]OLLILA S, MAKELA TP.The tumor suppressor kinase LKB1:lessons from mouse models[J].J Mol Cell Biol, 2011, 3 (6) :330-340.

[2]NAGARAJ AS, LAHTELA J, HEMMES A, et al.Cell of origin links histotype spectrum to immune microenvironment diversity in non-small-cell lung cancer driven by mutant kras and loss of lkb1[J].Cell Rep, 2017, 18 (3) :673-684.

[3]RHODES LV, TATE CR, HOANG VT, et al.Regulation of triplenegative breast cancer cell metastasis by the tumor-suppressor liver kinase B1[J].Oncogenesis, 2015, 4:e168.

[4]MA LG, BIAN SB, CUI JX, et al.LKB1 inhibits the proliferation of gastric cancer cells by suppressing the nuclear translocation of Yap and beta-catenin[J].Int J Mol Med, 2016, 37 (4) :1039-1048.

[5]MORTON JP, JAMIESON NB, KARIM SA, et al.LKB1 haploinsufficiency cooperates with Kras to promote pancreatic cancer through suppression of p21-dependent growth arrest[J].Gastroenterology, 2010, 139 (2) :586-597.e1-6.

[6]HUANG YH, HE B, LI P, et al.Expression and prognostic value of LKB1 in hepatocellular carcinoma[J].J Hepatopancreatobiliary Surg, 2013, 25 (6) :467-470. (in Chinese) 黄约翰, 何彬, 李鹏, 等.LKB1在肝细胞癌中的表达及其对预后的影响[J].肝胆胰外科杂志, 2013, 25 (6) :467-470.

[7]LI J, LIU J, LI P, et al.Loss of LKB1 disrupts breast epithelial cell polarity and promotes breast cancer metastasis and invasion[J].J Exp Clin Cancer Res, 2014, 33:70.

[8]JI H, RAMSEY MR, HAYES DN, et al.LKB1 modulates lung cancer differentiation and metastasis[J].Nature, 2007, 448 (7155) :807-810.

[9]ZHUANG ZG, DI GH, SHEN ZZ, et al.Enhanced expression of LKB1 in breast cancer cells attenuates angiogenesis, invasion, and metastatic potential[J].Mol Cancer Res, 2006, 4 (11) :843-849.

[10]SHEN Z, WEN XF, LAN F, et al.The tumor suppressor gene LKB1 is associated with prognosis in human breast carcinoma[J].Clin Cancer Res, 2002, 8 (7) :2085-2090.

[11]Ministry of Health of the People’s Republic of China.Diagnosis, management, and treatment of hepatocellular carcinoma (V2011) [J].J Clin Hepatol, 2011, 27 (11) :1141-1159. (in Chinese) 中华人民共和国卫生部.原发性肝癌诊疗规范 (2011年版) [J].临床肝胆病杂志, 2011, 27 (11) :1141-1159.

[12]ZHU PL, YIN C, FENG JL.Progress in comprehensive treatment of primary liver cancercomprehensive treatment of primary liver cancer[J].J Clin Hepatol, 2015, 31 (6) :965-968. (in Chinese) 祝普利, 尹超, 冯建龙.原发性肝癌综合治疗进展[J].临床肝胆病杂志, 2015, 31 (6) :965-968.

[13]SHAW RJ, BARDEESY N, MANNING BD, et al.The LKB1tumor suppressor negatively regulates m TOR signaling[J].Cancer Cell, 2004, 6 (1) :91-99.

[14]SANCHEZ-CESPEDES M, PARRELLA P, ESTELLER M, et al.Inactivation of LKB1/STK11 is a common event in adenocarcinomas of the lung[J].Cancer Res, 2002, 62 (13) :3659-3662.

[15]GAO Y, XIAO Q, MA H, et al.LKB1 inhibits lung cancer progression through lysyl oxidase and extracellular matrix remodeling[J].Proc Natl Acad Sci U S A, 2010, 107 (44) :18892-18897.

[16]JIANG S, CHEN R, YU J, et al.Clinical significance and role of LKB1 in gastric cancer[J].Mol Med Rep, 2016, 13 (1) :249-256.

[17]WANG J, ZHANG K, WANG J, et al.Underexpression of LKB1 tumor suppressor is associated with enhanced Wnt signaling and malignant characteristics of human intrahepatic cholangiocarcinoma[J].Oncotarget, 2015, 6 (22) :18905-18920.

[18]HUANG YH, CHEN ZK, HUANG KT, et al.Decreased expression of LKB1 correlates with poor prognosis in hepatocellular carcinoma patients undergoing hepatectomy[J].Asian Pac J Cancer Prev, 2013, 14 (3) :1985-1988.

[19]HE M, TAUSSIG MJ.Eukaryotic ribosome display with in situ DNArecovery[J].Nat Methods, 2007, 4 (3) :281-288.

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