[关键词]
[摘要]
目的:探讨转凝蛋白(transgelin,TAGLN)在结直肠癌(colorectal cancer,CRC)组织中的表达及其对SW480细胞增殖、迁移及侵袭的影响。方法: 选取郑州大学附属肿瘤医院2015年5月至2016年8月收治的97例CRC患者的癌及配对的癌旁组织标本,以及人CRC细胞系SW620、SW480、HCT116和正常结直肠黏膜细胞株FHC,用免疫组化染色法检测CRC组织中TAGLN的阳性表达率,并分析其表达水平与患者临床病理特征的关系。用qPCR法和WB法分别检测CRC细胞中TAGLN mRNA及蛋白的表达水平。采用脂质体法将si-TAGLN、si-Ctrl转染进SW480细胞,用CCK-8法、划痕愈合实验、Transwell小室法分别检测沉默TAGLN对SW480细胞增殖、迁移及侵袭的影响,用WB法检测EMT相关蛋白上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)和波形蛋白(vimentin)的表达水平。结果: TAGLN 在 CRC 组织中的阳性表达率明显高于癌旁组织(P<0.01),其表达水平与CRC患者的TNM分期、肿瘤分化程度和淋巴结转移相关联(P<0.05或P<0.01)。TAGLN mRNA及蛋白在SW480细胞中的表达水平显著高于FHC细胞(均P<0.01)。沉默TAGLN后,SW480细胞的增殖、迁移及侵袭能力均显著降低(均P<0.01),细胞中E-cadherin表达水平升高而N-cadherin和vimentin表达水平降低(均P<0.01)。结论: TAGLN在CRC组织和细胞中高表达,沉默TAGLN可抑制CRC细胞的增殖、迁移及侵袭能力,其在CRC的发生发展中起重要作用。
[Key word]
[Abstract]
Objective: To investigate the expression of transgelin (TAGLN) in colorectal cancer (CRC) tissues and its effect on the proliferation, migration and invasion of CRC SW480 cells. Methods: Surgically resected CRC tissues and corresponding paracancerous tissues of 97 CRC patients from May 2015 to August 2016 in the Affiliated Tumor Hospital of Zhengzhou University were collected; In addition, CRC cell lines SW620, SW480, HCT116 and normal colorectal mucosal cell line FHC were also collected for this study. Immunohistochemical staining was used to detect the expression of TAGLN in CRC tissues, and the correlation between TAGLN and patients’clinicopathological features was analyzed. Quantitative Real-time quantitative polymerase chain reaction (qPCR)and Western blotting (WB) were used to detect the mRNA and protein expressions of TAGLN in CRC cell lines. si-TAGLN and si-Ctrl were respectively transfected into SW480 cells by liposome transfection method. The effects of silencing TAGLN on the proliferation, migration and invasion of SW480 cells were detected by CCK-8, Wound-healing assay and Transwell assay,respectively; and the expression of EMT-related proteins E-cadherin, N-cadherin and vimentin were detected by WB. Results: The positive expression rate of TAGLN in CRC tissues was significantly higher than that in para-cancerous tissues (P<0.01), and TAGLN expression was correlated with TNM stage, degree of tumor differentiation and lymph node metastasis in CRC patients (P<0.05 or P<0.01). The mRNA and protein expression levels of TAGLN in SW480 cells were significantly higher than those in FHC cells (all P<0.01). After TAGLN silence, the proliferation, invasion and migration ability of SW480 cells were significantly reduced (all P<0.01), the expression level of E-cadherin in SW480 cells was increased, while the expression levels of N-cadherin and vimentin were decreased (all P<0.01). Conclusion: TAGLN is highly expressed in CRC tissues and cells. Silencing TAGLN can inhibit the proliferation, invasion and migration of CRC cells, suggesting that TAGLN plays an important role in the occurrence and development of CRC.
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[基金项目]
河南省医学科技攻关计划资助项目(No. 201702246)