摘要
目的研究遗传印记基因PEG10蛋白在大肠癌细胞中的亚细胞定位,探讨PEG10在大肠癌侵袭、生长中的作用机制。方法应用激光扫描共聚焦显微镜成像方法检测PEG10蛋白在大肠癌细胞中的亚细胞定位;构建siPEG10干扰质粒,高效稳定转染至大肠癌细胞系SW620,采用嘌呤霉素对干扰质粒细胞株进行筛选;利用Westernblot法检测PEG10基因在蛋白水平的表达情况;通过Transwell小室技术探讨PEG10对SW620细胞侵袭和生长的影响;分组构建动物模型,把PEG10-siRNA后的SW620细胞在C57BL/6小鼠皮下进行注射,观察其移植瘤的增长情况。结果激光扫描共聚焦显微镜成像显示PEG10主要定位在细胞质;成功构建了PEG10的干扰质粒;Western-blot方法验证了细胞株中PEG10基因表达有效的被抑制;Transwell小室实验显示干扰PEG10后大肠癌细胞SW620侵袭能力减弱;在体内实验中,注射干扰PEG10后大肠癌细胞SW620的小鼠皮下瘤体较对照组小。结论 PEG10在体内外对小鼠大肠癌细胞SW620的侵袭和生长能力有促进作用。
Objective To study the subcellular localization of PEG10 gene in colorectal cancer cells, and explore the role of PEG10 in the invasion and growth of colorectal cancer. Methods Subcellular localization of PEG10 protein was detected by laser scanning confocal microscopy imaging. PEG10siRNA expression vector was constructed and stably transfected into SW620 cells. The expression of PEG10 gene was detected by Westernblot. The effect of PEG10 on the invasion and growth of SW620 cells was investigate with Transwell assay. Observing The tumor growth was observed when the SW620 cells after PEG10siRNA was injected in C57BL/6 mice. Results Laser scanning confocal microscopy imaging showed that PEG10 was mainly located in the cytoplasm. The interference plasmid PEG10 was successfully constructed. Transwell cell experiments indicated that siPEG10 inhibited colorectal cancer cell SW620 invasion ability. In vivo, siPEG10 SW620 cells were injected into subcutaneous tumor inhibited colorectal cancer cells growth compared with the control group. Conclusion PEG10 can promoted the invasion and growth of colorectal cancer cell lines in vitro and in vivo.
出处
《西部医学》
2018年第1期16-20,共5页
Medical Journal of West China
基金
厦门市科技计划项目(3502Z20144035)
