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@#[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 para-cancerous 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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@#[摘 要] 目的：探讨敲减中心体相关激酶2（never in mitosis A-related kinase 2，NEK2）对结直肠癌细胞5-FU化疗敏感性的影响及其可能的机制。方法：采用qPCR和Western blotting检测结直肠癌细胞中NEK2 mRNA及蛋白的表达水平。构建针对NEK2基因的小干扰RNA（siRNA）并转染至结直肠癌细胞HCT116及SW620，实验分为阳性干扰组1（转染NEK2 siRNA1）、阳性干扰组2（转染NEK2 siRNA2）和阴性对照组（转染si-NC），均用5-FU处理。采用CCK-8实验、V-FICT/PI Annexin双染色流式细胞术实验观察敲减NEK2基因对5-FU作用下结肠癌细胞的增殖、周期分布及凋亡的影响，采用Western blotting检测敲减NEK2基因对5-FU作用下结直肠癌细胞内Wnt/β-catenin信号通路相关蛋白表达的影响。结果：NEK2蛋白及mRNA在结直肠癌细胞HCT116、SW620中均呈高表达（P<0.05或P<0.01），转染NEK2 siRNA可高效抑制HCT116、SW620细胞中NEK2蛋白及mRNA表达（均P<0.01）。经不同浓度5-FU作用后，阳性干扰组1和阳性干扰组2的细胞存活率和IC50均显著低于阴性对照组（均P<0.01），细胞发生G0/G1期阻滞且凋亡率显著升高（均P<0.01），胞核β-catenin、c-myc和cyclin D1表达水平显著下降而胞质β-catenin表达水平升高（均P<0.01）。结论：敲减NEK2基因可有效提高人结直肠癌细胞对5-FU的化疗敏感性，该作用可能是通过调控Wnt/β-catenin信号通路相关蛋白表达来实现的。
ABSTRACT
@# Objective: To explore the expression of IQGAP1 (Ras GTPase-activating-like protein containing IQ domain) in esophageal squamous cell carcinoma (ESCC) tissues and cell lines and its effect on the proliferation and invasion of TE-2 cell. Methods: Totally 125 pairs of cancer tissues and para-cancerous tissues from ESCC patients, who underwent surgical resection inAffiliated Tumor Hospital of Zhengzhou University from January 2015 to December 2016, were included in this study; in addition, ESCC cell lines (TE-2, TE3, ECA109) and normal esophageal epithelial cell line Het-1A were also collected. The expression of IQGAP1 was detected by immunohistochemical staining and its relationship with cliniopathological features was also analyzed. IQGAP1 mRNA and protein expressions in ESCC cell lines were detected by Real-time quantitative PCR (qPCR) and Western blotting, respectively. TE-2 cells were transfected with si-IQGAP1 (positive transfection group) and si-CTRL (negative control group) plasmids, and the effects of IQGAP1 silencing on the proliferation and invasion of TE-2 cells were detected by MTT and Transwell assay. The expressions of E-cadherin and Ncadherin were detected by Western blotting. Results: The positive expression rate of IQGAP1 in ESCC tissues was significantly higher than that in para-cancerous tissues (P<0.05), which was closely related to tumor stage and histologic grade (all P<0.05). The mRNAand protein expressions of IQGAP1 in TE-2, TE-3 and ECA109 cells were significantly higher than those in Het-1Acells (all P<0.05).After IQGAP1 was silenced, compared with the negative control group and the blank group, the expression of IQGAP1 mRNAand protein in the positive transfection group significantly decreased (all P<0.05); the proliferation and invasiveness of TE-2 cells significantly decreased (all P<0.05); E-cardherin was up-regulated while N-cardherin was down-regulated (all P<0.05) in the positive interference group. Conclusion: IQGAP1 is highly expressed in ESCC tissues, and si-IQGAP1 can inhibit the proliferation and invasion of TE-2 cells, which plays an important role in the occurrence and development of ESCC.