Department of Interventional Radiology, Shanghai 10th People's Hospital, Tongji University, No. 301, Yanchang Road, Shanghai, 200072, China.
Mol Biol Rep. 2013 Feb;40(2):949-56. doi: 10.1007/s11033-012-2136-4. Epub 2012 Oct 14.
This study intended to investigate the expression of the ZEB1 and E-cadherin proteins in lung squamous cell carcinoma (LSCC) tissues and to examine the clinicopathological correlation between protein levels and LSCC. RT-PCR and Western blot were used to examine the expression of ZEB1 and E-cadherin mRNAs and proteins in LSCC tissues as well as in adjacent normal tissues, and then analyze the relationship between the clinicopathological characteristics and the expression changes of ZEB1 and E-cadherin mRNAs in LSCC. In addition, RNAi was used to knockdown the expression of the ZEB1 gene in Human HCC827 cells; subsequently, changes in the invasive ability of the resultant cells were studied. The positive rates of ZEB1 and E-cadherin mRNAs in LSCC tissues were 69.2 and 38.5 %, respectively. They differed significantly from the corresponding positive rates in the adjacent normal lung tissues (15.4 and 80.8 %, p < 0.05). There was a negative correlation between the protein levels of ZEB1 and E-cadherin in LSCC tissues (r = -0.714, p < 0.001); in addition, it was found that ZEB1 protein expression in LSCC tissues was significantly higher than that in the neighboring normal lung tissues (p < 0.05), and its expression was also significantly higher in patients with lymph node metastases and distant metastases compared to those patients without metastatic disease (p < 0.05). On the contrary, E-cadherin expression was significantly lower in LSCC tissues than that in the neighboring normal tissue (p < 0.05). It was lower in patients with lymph node metastasis and distant metastasis compared to patients without metastatic disease (p < 0.05). However, the expression of ZEB1 and E-cadherin was independent of gender, age, tumor size, or tumor differentiation level (p > 0.05). Transfection of ZEB1 siRNA into HCC827 cells significantly reduced the ZEB1 protein level (p < 0.01) and significantly elevated E-cadherin levels (p < 0.01). Moreover, significantly less ZEB1 siRNA-transfected cells migrated through Transwell chambers in the LSCC tissue than that in the control groups (untransfected or transfected with control siRNA, p < 0.01). The expression of the ZEB1 gene in LSCC tissues is downregulated with the expression of E-cadherin. On the other hand, the expression of siRNA against ZEB1 promotes E-cadherin expression and suppresses the invasive ability conferred by E-cadherin. In conclusion, our data suggested that overexpression of the ZEB1 gene is possibly associated with the occurrence, development, invasion of LSCC.
本研究旨在探讨 ZEB1 和 E-钙黏蛋白蛋白在肺鳞癌(LSCC)组织中的表达,并分析其蛋白水平与 LSCC 临床病理特征之间的相关性。采用 RT-PCR 和 Western blot 检测 LSCC 组织及相邻正常组织中 ZEB1 和 E-钙黏蛋白 mRNA 和蛋白的表达情况,并分析 ZEB1 和 E-钙黏蛋白 mRNA 在 LSCC 中的表达变化与临床病理特征之间的关系。此外,利用 RNAi 技术敲低人 HCC827 细胞中 ZEB1 基因的表达,研究由此导致的细胞侵袭能力的变化。LSCC 组织中 ZEB1 和 E-钙黏蛋白 mRNA 的阳性率分别为 69.2%和 38.5%,显著低于相应的癌旁正常肺组织(15.4%和 80.8%,p<0.05)。LSCC 组织中 ZEB1 和 E-钙黏蛋白蛋白的表达水平呈负相关(r=-0.714,p<0.001);此外,LSCC 组织中 ZEB1 蛋白的表达明显高于癌旁正常肺组织(p<0.05),且淋巴结转移和远处转移患者的表达明显高于无转移疾病患者(p<0.05)。相反,LSCC 组织中 E-钙黏蛋白的表达明显低于癌旁正常组织(p<0.05),且淋巴结转移和远处转移患者的表达明显低于无转移疾病患者(p<0.05)。然而,ZEB1 和 E-钙黏蛋白的表达与性别、年龄、肿瘤大小或肿瘤分化程度无关(p>0.05)。转染 ZEB1 siRNA 显著降低 HCC827 细胞中的 ZEB1 蛋白水平(p<0.01),并显著升高 E-钙黏蛋白水平(p<0.01)。此外,转染 ZEB1 siRNA 的细胞穿过 LSCC 组织 Transwell 小室的迁移明显少于对照组(未转染或转染对照 siRNA,p<0.01)。LSCC 组织中 ZEB1 基因的表达下调伴随着 E-钙黏蛋白的表达。另一方面,针对 ZEB1 的 siRNA 的表达促进了 E-钙黏蛋白的表达,并抑制了 E-钙黏蛋白赋予的侵袭能力。综上所述,我们的数据表明 ZEB1 基因的过表达可能与 LSCC 的发生、发展、侵袭有关。