H2-calponin在食管鳞癌中的表达及意义

周玲1, 凌锐1, 周月鹏1, 毛朝明2, 陈德玉1

江苏大学学报(医学版) ›› 2018, Vol. 28 ›› Issue (02) : 104-108,116.

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江苏大学学报(医学版) ›› 2018, Vol. 28 ›› Issue (02) : 104-108,116.
基础医学

H2-calponin在食管鳞癌中的表达及意义

作者信息 +

Expression of H2-calponin in esophageal squamous cell carcinoma and its clinical significance

  • ZHOU Ling1, LING Rui1, ZHOU Yue-peng1, MAO Chao-ming2, CHEN De-yu1
Author information +

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。

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.

关键词

H2-calponin / NF-κB / 食管鳞癌

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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[J]. Journal of Jiangsu University(Medicine Edition), 2018, 28(02): 104-108,116

参考文献

[1]Liu R, Jin JP. Calponin isoforms CNN1, CNN2 and CNN3: Regulators for actin cytoskeleton functions in smooth muscle and nonmuscle cellsJ. Gene, 2016, 585(1):143-153.

2]Jiang WR, Cady G, Hossain MM, et al. Mechanoregulation of H2calponin gene expression and the role of Notch signalingJ. J Biol Chem, 2014, 289(3):1617-1628.
 

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