天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (3): 225-229.doi: 10.11958/20171068

• 细胞与分子生物学 •    下一篇

慢病毒介导的PRL-3 shRNA对结肠癌细胞增殖、侵袭、凋亡的影响

王燕,张姣，王会峰，王宁菊   

  1. 宁夏医科大学总医院肿瘤内科（邮编750004）
  • 收稿日期:2017-10-11 修回日期:2018-01-03 出版日期:2018-03-15 发布日期:2018-03-23
  • 通讯作者: 王燕 E-mail:mgwy1974@163.com
  • 基金资助:
    宁夏自然科学基金项目

Effects of shRNA targeting PRL-3 on proliferation, invasion and apoptosis of human colorectal cancer cells

WANG Yan, ZHANG Jiao, WANG Hui-feng, WANG Ning-ju   

  1. Department of Medical Oncology, General Hospital of Ningxia Medical University, Yinchuan 750004, China
  • Received:2017-10-11 Revised:2018-01-03 Published:2018-03-15 Online:2018-03-23

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摘要： 目的 观察慢病毒介导的 shRNA 沉默肝再生磷酸酶-3（PRL-3）基因对结肠癌 SW480 细胞增殖、侵袭、凋亡的影响。方法 实验分组为空白对照组、阴性对照组、转染组。将携带 PRL-3 shRNA 的慢病毒载体转染结肠癌SW480 细胞，建立稳定沉默 PRL-3 的细胞株，real-time PCR 检测转染后 PRL-3 mRNA 的相对表达水平。采用 MTT法、平板克隆形成实验检测转染后细胞增殖能力；采用 Transwell 侵袭实验、侵袭小室法检测转染后细胞迁移及侵袭能力；采用流式细胞术检测转染后细胞凋亡率变化。结果 稳定沉默 PRL-3 的细胞株构建成功，转染组 PRL-3mRNA 的相对表达水平低于空白对照组、阴性对照组（P＜0.05），空白对照组、阴性对照组比较差异无统计学意义。PRL-3 shRNA 转染 SW480 细胞 72 h 后，转染组与空白对照组、阴性对照组比较，细胞增殖能力受到抑制，转染 120 h时最明显（P＜0.05）。转染组克隆形成能力较空白对照组、阴性对照组下降（P＜0.05）。转染组与空白对照组、阴性对照组比较，细胞迁移、侵袭能力下降，凋亡率增加（P＜0.05）。结论 结肠癌 SW480 细胞转染 PRL-3 shRNA 可减少 PRL-3 的表达，有效抑制 SW480 细胞增殖，促进其凋亡，PRL-3 可能成为治疗结肠癌的靶基因。

关键词: 基因沉默, 结肠肿瘤, 细胞系, 肿瘤, 细胞增殖, 细胞凋亡, 肝再生磷酸酶-3

Abstract: Objective To investigate the effects of PRL-3 shRNA mediated by lentivirus on proliferation, invasion and apoptosis of human colorectal cancer SW480 cells. Methods There were three experimental groups in this study, which included blank control group, negative control group and transfected group. Colorectal cancer SW480 cells were transfected with lentiviral interference vector carrying PRL-3 shRNA to build a stable PRL-3-silencing cell line. The expression of PRL-3 mRNA was detected by real-time quantitative polymerase chain reaction (real-time PCR). Cell proliferation was analyzed by MTT method and colony formation assay. Invasion and migration were measured by Transwell assay and invasion chamber. Apoptosis rate was performed by flow cytometry. Results The stable PRL-3-silencing cell line was successfully constructed. Compared with the blank control group and negative control group, the relative expression levels of PRL-3 mRNA were reduced in transfected group after transfection with PRL-3 shRNA (P＜0.05), but there was no significant difference between the blank control group and the negative control group. After transfection with PRL-3 shRNA for 72 h, the proliferation of SW480 cells was significantly lower in transfected group than that of the blank control group and the negative control group, and the proliferation decreased significantly in 120 h (P＜0.05).Compared with the blank control group and negative control group, the ability of colony formation was also weakened in the transfected group (P＜0.05).Compared with the blank control group and negative control group, the migration and invasion ability were decreased in the transfected group (P＜0.05), and the apoptosis rate was increased in the transfected group (P＜0.05). Conclusion PRL-3 shRNA can inhibit the expression of PRL-3 and the proliferation, promote the apoptosis of SW480, which indicates that PRL-3 may become a target for colorectal carcinoma treatment.

Key words: gene silencing, colonic neoplasms, cell line, tumor, cell proliferation, apoptosis, phosphatase of regenerating liver-3

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