阻断CLC-3氯通道对结直肠癌细胞株生存率和侵袭转移能力的影响及其分子机制

Blockade of CLC-3 chloride channel inhibited the viability and invasion of colorectal cancer cells

目的:通过研究CLC-3在结直肠组织中的表达,探讨CLC-3对结直肠癌细胞株SW480、SW620的细胞生存率、侵袭转移能力的影响及其潜在的机制。方法:采用RT-PCR方法测定不同分期结直肠癌组织和正常结直肠组织中CLC-3的mRNA表达水平。运用CLC-3阻断剂DIDS、NPPB阻断SW480、SW620细胞的CLC-3表达后,采用CCK-8法实验检测细胞生存率,细胞侵袭实验检测细胞侵袭转移情况;并采用免疫印迹法测定阻断CLC-3表达后SW480、SW620细胞Wnt/β-catenin信号通路相关蛋白表达。结果:结直肠癌组织中CLC-3 mRNA表达水平高于结直肠炎黏膜组织和正常结直肠组织(P<0.05),且CLC-3 mRNA表达和结直肠分期相关。抑制CLC-3表达后,SW480、SW620细胞的生存率和侵袭转移能力降低(P<0.05),且β-catenin、C-myc、cyclin D1、Ki-67、survivin表达降低(P<0.05)。结论:CLC-3高表达与结直肠的发生发展有潜在联系,其机制可能与Wnt/β-catenin有关,为CLC-3作为治疗结直肠癌的潜在新靶点提供实验基础。

To examine the expression of CLC-3 in colorectal tissues and the effect of CLC-3 on the viability and invasion of colorectal cancer（CRC） SW480 and SW620 cells. Methods： The m RNA levels of CLC-3 in CRC cell lines were determined by RT-PCR. CLC-3 expression was inhibited by adding DIDS or NPPB to the CRC cells. Subsequently, cell viability and invasion were assessed by CCK-8 assay and Transwell assay, respectively. In addition, the effects of DIDS and NPPB on the Wnt or β-catenin signaling pathways were determined by Western blot analysis. Results： The m RNA level of CLC-3 was remarkably increased in the CRC tissues compared with that in normal colorectal tissues（P〈0.05） and was positively correlated with the T stage of CRC. The blockade of CLC-3 inhibited the viability and invasion of CRC cells（P〈0.05）. The expression of β-catenin, C-myc, cyclin D1, Ki-67, and survivin were evidently reduced by the inhibition of CLC-3（P〈0.05）. Conclusion： The inhibition of CLC-3 decreases the cell viability and invasion of CRC cells by reducing the expression of the proteins related to the Wnt or β-catenin signaling pathway.