恶性黑素瘤,TGB8,Wnt/β-catenin信号通路,侵袭,转移 ," /> 恶性黑素瘤,TGB8,Wnt/β-catenin信号通路,侵袭,转移 ,"/> <p class="MsoNormal"> <span>整合素</span>β<span>8</span><span>在恶性黑素瘤细胞增殖及侵袭中的作用</span>

中国麻风皮肤病杂志 ›› 2017, Vol. 33 ›› Issue (2): 83-87.

• 论著 • 上一篇    下一篇

整合素β8在恶性黑素瘤细胞增殖及侵袭中的作用

廖晓容1  高永良2   

  1. 1重庆市第六人民医院皮肤科，重庆，400060

    2重庆医科大学附属第一医院皮肤科，400016

  • 出版日期:2017-02-15 发布日期:2018-12-13
  • 通讯作者: 高永良，E-mail：gao2418@126.com

The role of integrin β8 in the invasion and metastasis of malignant melanoma

LIAO Xiaorong1, GAO Yongliang2   

  1. 1. Department of Dermatology, the Sixth People's Hospital of Chongqing, Chongqing 400060, China; 2. Department of Dermatology, the First  Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China

  • Online:2017-02-15 Published:2018-12-13
  • Contact: GAO Yongliang, E-mail: gao2418@126.com

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摘要：

目的：明确整合素β8（ITGB8）在恶性黑素瘤侵袭转移中的作用。

方法：Transwell侵袭实验在黑素瘤细胞系A375中获得侵袭性癌细胞和相对静止性癌细胞，RT-PCR比较两者整合素家族的表达。流式分选ITGB8细胞，比较其侵袭能力的差异。使用siRNA敲除技术敲低ITGB8，敲低效率分别为：si1组 50%，si2组70%，si3组80%，比较其增殖、侵袭能力的变化。Western bolt检测细胞周期蛋白、基质金属蛋白酶及EMT相关分子的表达变化，提取细胞核蛋白比较细胞核内β-catenin的变化。利用TCF4的启动子报告质粒检测Wnt/β-catenin信号通路的激活情况。结果：ITGB8在侵袭性癌细胞中高表达，是静止性癌细胞的5倍（P<0.01）；ITGB8阳性组侵袭细胞数为516±45，阴性组为120±22，差异有显著性（P<0.01）； 与 Mock组细胞相比，si2和si3组细胞的增殖速度明显减弱（P<0.01），敲低后细胞阻滞在S期。与Mock细胞相比Cyclin D1、MMP2、MMP9、MMP7、Ncadherin、 Vimentin在si2和si3细胞中的表达明显减少，Ecadherin明显增加。si2、si3细胞核内β-catenin相比Mock组明显减少（P<0.05）。敲低ITBG8后TCF4的启动子活性明显降低。结论：ITGB8对恶性黑素瘤的增殖及侵袭转移具有重要作用，可能是通过调控Wnt/β-catenin信号通路发挥作用。

关键词: 恶性黑素瘤')">

恶性黑素瘤, TGB8, Wnt/β-catenin信号通路, 侵袭, 转移

Abstract:

Objective: To determine the role of integrin β8 (ITGB8) in the invasion and proliferation of malignant melanoma. Methods: Melanoma cell line A375 cancer cells were divided into invasive cancer cells and non-invasive cancer cells through Transwell invasion experiment. The expression of integrin family in the A375 cancer cells was detected by RT-PCR. The invasive abilities of the A375 cancer cells of ITGB8 negative and positive cells were compared by flow cytometer. The ITGB8 positive A375 cancer cells were knocked out by siRNA knockout technology and the removal efficiency was 50% in si1 group, 70% in si2 group and 80% in si3 group. The proliferation and invasion were compared among the three groups. The cell cycle protein molecules, matrix metalloproteinases and EMT related molecule were detected by Western bolt. The nucleus protein was extracted to compare the changes of Wnt/beta-catenin in the nucleus of the A375 cancer cells in different knockout efficiency. The activation of beta-catenin signaling pathway was detected through TCF4 promoter report plasmid (TOP/FOP) test. Results: The expression level of ITGB8 in invasive cancer cells was 5 times as high as in non-invasive β8 (P<0.01). The number of ITGB8 positive invasive cancer cells was 516±45, which was higher than that in ITGB8 negative invasive cancer cells (120±22), with a significant difference (P<0.01). The proliferation rate of the cells in si2 group and si3 group was lower than that in MOCK cells. Compared with the cells in MOCK group, the expression of Cyclin D1, MMP2, MMP9, MMP7 and Ncadherin, Vimentin decreased and Ecadherin increased in si2 and si3 groups. The amount of the beta-catenin in cell nucleus in the cells of si2 and si3 group was lower than that in Mock group. Promoter activity of TCF4 decreased after ITBG8 removed. Conclusion: ITGB8 plays an important role in the process of invasion and proliferation of malignant melanoma, which may through regulating the Wnt/beta-catenin signaling pathways.

体" >?q?<?h ??c nt face="Calibri" >ITGB8阳性组侵袭细胞数为516±45，阴性组为120±22，差异有显著性（P<0.01）； 与 Mock组细胞相比，si2和si3组细胞的增殖速度明显减弱（P<0.01），敲低后细胞阻滞在S期。与Mock细胞相比Cyclin D1、MMP2、MMP9、MMP7、Ncadherin、 Vimentin在si2和si3细胞中的表达明显减少，Ecadherin明显增加。si2、si3细胞核内β-catenin相比Mock组明显减少（P<0.05）。敲低ITBG8后TCF4的启动子活性明显降低。结论：ITGB8对恶性黑素瘤的增殖及侵袭转移具有重要作用，可能是通过调控Wnt/β-catenin信号通路发挥作用。