Lu X, Zhou C, Li R F, Ye J W, Zhai W L
Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital of Zhengzhou University, Digestive Organ Transplantation of Henan Province, Zhengzhou 450052, China.
Zhonghua Wai Ke Za Zhi. 2018 Aug 1;56(8):617-622. doi: 10.3760/cma.j.issn.0529-5815.2018.08.013.
To investigate the effects of Kindlin-2 on malignant phenotypes of human gallbladder cancer cells and discuss the mechanisms. The expression level of Kindlin-2 in 30 cases of gallbladder cancer tissues and adjacent non-tumoral tissues collected from the First Affiliated Hospital of Zhengzhou University between September 2012 and May 2013 was assessed by real-time PCR and immunohistochemistry.Lentivirus-mediated Kindlin-2 overexpression was used in gallbladder cancer cell lines GBC-SD and SGC-996.Transwell assay and adhesion assay were investigated to explore the functional role of Kindlin-2 on gallbladder cancer cells.Western Blot was used to test the protein change of epithelial-mesenchymal transition(EMT) characteristics. The -test was used to analyzed results. The RNA and protein levels of Kindlin-2 in gallbladder cancer tissues were higher than in the non-tumoral tissues (=4.372, =0.001; =7.477, =0.000). The expression level of Kindlin-2 in gallbladder cancer tissues was correlated with Nevin stage(χ(2)=5.932, =0.035). Compared with control groups, the cell-matrix adhesion ability of GBC-SD and SGC-996 with Kindlin-2 overexpression was obviously promoted(1.66±0.03 . 1.07±0.22, =2.710, =0.041; 2.66±0.24 . 1.03±0.02, =6.610, =0.020). The number of GBC-SD and SGC-996 cells with Kindlin-2 overexpression passing through the Transwell chamber matrix increased significantly compared with the control groups(116.1±13.9 . 54.7±8.4, =3.781, =0.019; 136.3±7.5 . 64.3±6.4, =7.302, =0.002). The wound healing rate of GBC-SD with Kindlin-2 overexpression at 12-hour and 24-hour was higher than that of the group ((42.9±2.2)% . (29.7±1.7)%, =4.690, =0.009; (65.0±2.4)% .(40.4±2.0)%, =7.945, =0.001). The wound healing rate of SGC-996 with Kindlin-2 overexpression at 12-hour and 24-hour was also higher than that of the group ((32.9±1.3)% . (24.1±1.5)%, =4.518, =0.011; (51.3±1.1)% . (39.2±1.1)%, =8.001, =0.001). The characteristics of EMT were induced in gallbladder cancer cells with Kindlin-2 overexpression, including the up-regulation of N-cadherin, Vemintin and the down-regulation of E-cadherin. The expression of Kindlin-2 is up-regulated in gallbladder cancer tissues and Kindlin-2 promoted the malignant phenotypes of gallbladder cancer cells partially by epithelial-mesenchymal transition.
探讨Kindlin-2对人胆囊癌细胞恶性表型的影响并探讨其机制。采用实时荧光定量PCR和免疫组织化学方法检测2012年9月至2013年5月郑州大学第一附属医院收集的30例胆囊癌组织及癌旁非肿瘤组织中Kindlin-2的表达水平。采用慢病毒介导的Kindlin-2过表达技术处理胆囊癌细胞系GBC-SD和SGC-996。通过Transwell实验和黏附实验探讨Kindlin-2对胆囊癌细胞的功能作用。采用蛋白质免疫印迹法检测上皮-间质转化(EMT)相关蛋白的变化。采用t检验分析结果。胆囊癌组织中Kindlin-2的RNA和蛋白水平均高于癌旁非肿瘤组织(t=4.372,P=0.001;t=7.477,P=0.000)。Kindlin-2在胆囊癌组织中的表达水平与Nevin分期相关(χ²=5.932,P=0.035)。与对照组相比,过表达Kindlin-2的GBC-SD和SGC-996细胞与基质的黏附能力明显增强(1.66±0.03比1.07±0.22,t=2.710,P=0.041;2.66±0.24比1.03±0.02,t=6.610,P=0.020)。过表达Kindlin-2的GBC-SD和SGC-996细胞穿过Transwell小室基质的数量较对照组明显增加(116.1±13.9比54.7±8.4,t=3.781,P=0.019;136.3±7.5比64.3±6.4,t=7.302,P=0.002)。过表达Kindlin-2的GBC-SD在12小时和24小时的伤口愈合率高于对照组((42.9±2.2)%比(29.7±1.7)%,t=4.690,P=0.009;(65.0±2.4)%比(40.4±2.0)%,t=7.945,P=0.001)。过表达Kindlin-2的SGC-996在12小时和24小时的伤口愈合率也高于对照组((32.9±1.3)%比(24.1±1.5)%,t=4.518,P=0.011;(51.3±1.1)%比(39.2±1.1)%,t=8.001,P=0.001)。过表达Kindlin-2可诱导胆囊癌细胞发生EMT,表现为N-钙黏蛋白、波形蛋白表达上调,E-钙黏蛋白表达下调。Kindlin-2在胆囊癌组织中表达上调,其可能通过部分诱导上皮-间质转化促进胆囊癌细胞的恶性表型。