H2-calponin在食管鳞癌中的表达及意义

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摘要目的: 探讨H2钙调理蛋白(H2calponin)在食管鳞癌中的表达及意义。方法: 免疫组化技术检测30例食管鳞癌患者癌组织及癌旁组织芯片中H2calponin的表达水平；体外培养人食管鳞癌ECA109、TE1细胞株和正常食管上皮Het1a细胞；荧光定量PCR和蛋白质印迹法检测食管鳞癌细胞株中H2calponin的表达；将食管鳞癌TE1细胞分为空白对照组（常规培养）、阴性对照组（转染空载体siRNA）和H2calponin siRNA组（转染H2calponin siRNA），采用蛋白质印迹法测定细胞中增殖细胞核抗原(PCNA)、E钙黏蛋白(Ecadherin)，波形蛋白以及基质金属蛋白酶2(MMP2)、MMP9、内皮生长因子A(VEGFA)、血管内皮生长因子C(VEGFC)蛋白的表达；ELISA法测定各组细胞培养上清液中MMP2，MMP9和VEGFC含量；荧光定量PCR检测NFκB通路抑制剂PDTC处理前（0 h）及处理后0.5, 1, 2, 4, 8, 12 h H2calponin mRNA的表达；蛋白质印迹技术检测H2calponin及NFκB通路蛋白的表达。结果：免疫芯片结果显示，食管鳞癌组织中H2calponin表达量低于癌旁组织， H2calponin阳性表达率与食管鳞癌患者性别、年龄、肿瘤部位及病理分级之间无明显相关性(P均>0.05)；荧光定量PCR和蛋白质印迹结果显示，食管鳞癌细胞中H2calponin表达明显低于正常食管细胞(P<0.01)；下调H2calponin表达后，PCNA、E钙黏蛋白和VEGFA表达无明显变化，而波形蛋白、MMP2、MMP9和VEGFC表达明显增加；ELISA结果显示，MMP2、MMP9和VEGFC分泌量明显增加(P<0.05)；PDTC处理后可显著增加IκBα的蛋白表达进而恢复并上调H2calponin的表达(P<0.01)。结论： NFκB通路通过抑制H2calponin表达保证波形蛋白，MMP2，MMP9，VEGFC持续表达，促进食管鳞癌细胞侵袭、转移发生。

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关键词 ： H2-calponin, NF-κB, 食管鳞癌

Abstract：Objective: To investigate the expression and significance of H2calponin in esophageal squamous cell carcinoma(ESCC). Methods: The expression of H2calponin in a tissue microarray consisted of 30 patients with ESCC and adjacent tumor tissues were detected by immunohistochemistry. The expression of H2calponin in ESCC cell line (ECA109 and TE1) and normal human esophageal epithelial cell(Het1a) were detected by RTPCR and Western blotting. TE1 cells were divided into blank control (routinely cultured), negative control (transfected with negative siRNA) and siRNAH2calponin group (transfected with H2calponin siRNA). The expression of PCNA, Ecadherin, Vimentin, MMP2, MMP9, VEGFA and VEGFC in each group was detected by Western blotting. The secretion of MMP2, MMP9 and VEGFC were detected by ELISA. The expression changes of H2calponin before(0 h) and after PDTC treatment (0.5, 1, 2, 4, 8, 12 h) were detected by RTPCR. Western blotting was used to determine the expresion of H2calponin and NFκB pathway related protein. Results: H2calponin were weakly expressed in ESCC tissues compared with adjacent normal tissues; expression of H2calponin was not significantly correlated with gender, age, tumor location and pathological grade in esophageal squamous cell carcinoma(P>0.05). ESCC cell lines showed higher H2calponin expression levels than that in Het1a cells(P<0.01). There were no significant change of PCNA, Ecadherin and VEGFA expression, while the expression of MMP2, MMP9 and VEGFC were upregulated after siRNAH2calponin treatment(P<0.05). PDTC treatment could rapidly restored the expression of H2calponin(P<0.01) by increasing IκBα expression. Conclusion: NFκB pathway promotes the invasion and metastasis of ESCC by restraining H2calponin through enhancing continuous expression of Vimentin, MMP2, MMP9 and VEGFC.

收稿日期: 2017-10-23

通讯作者: 陈德玉(通讯作者)，主任医师，教授，博士生导师，E-mail:cdeyu@hotmail.com

引用本文:

周玲1，凌锐1，周月鹏1，毛朝明2，陈德玉1. H2-calponin在食管鳞癌中的表达及意义[J]. 江苏大学学报:医学版, 2018, 28(02): 104-108,116. ZHOU Ling1, LING Rui1, ZHOU Yue-peng1, MAO Chao-ming2, CHEN De-yu1. Expression of H2-calponin in esophageal squamous cell carcinoma and its clinical significance. Journal of Jiangsu University(Medicine Edition), 2018, 28(02): 104-108,116.

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http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2018/V28/I02/104