LRP16调控EGFR通路对结肠癌细胞增殖、迁移的影响

张燕萍; 张琳; 王振; 李鑫; 汪静宇

doi:10.3969/j.issn.1671-038X.2021.08.08

LRP16调控EGFR通路对结肠癌细胞增殖、迁移的影响

- 嘉兴市第一医院病理科(浙江嘉兴, 314000)
基金项目:
嘉兴市科技计划项目（No：2019AY32026）

详细信息

通讯作者: 汪静宇,E-mail:11018140@zju.edu.cn

中图分类号: R735.3

收稿日期: 2021-03-29

Effect of LRP16 regulating EGFR pathway on proliferation and migration of colon cancer cells

- Department of Pathology, Jiaxing First Hospital, Jiaxing, 314000, China

More Information

Corresponding author: WANG Jingyu, E-mail: 11018140@zju.edu.cn

Received Date: 29 March 2021

摘要

摘要: 目的：研究人白血病相关蛋白16（LRP16）调控表皮生长因子受体（EGF/EGFR）通路对结肠癌细胞生物学行为的影响。方法：体外培养结肠癌SW620细胞株，分为阴性对照组（siRNA-NC组）、沉默LRP16表达组（LRP16-siRNA组）、外源性表皮生长因子（EGF）组（LRP16-siRNA+重组人EGF组）。采用实时荧光定量PCR （qRT-PCR）法检测各组SW620细胞LRP16 mRNA水平；CCK-8法、平板克隆实验分别检测各组SW620细胞增殖情况、克隆形成率；transwell实验检测各组SW620细胞迁移能力；免疫印迹法检测增殖细胞核抗原（PCNA）、迁移侵袭相关蛋白[E-钙黏附蛋白（E-cadherin）、N-钙黏附蛋白（N-cadherin）]及通路相关蛋白[EGFR、大鼠肉瘤（Ras）、Raf]蛋白表达情况。结果：细胞LRP16转染成功；与siRNA-NC组比较，LRP16-siRNA组SW620细胞增殖抑制率、E-Cadherin蛋白表达显著升高（P<0.05），克隆形成率、迁移细胞数、LRP16、PCNA、N-Cadherin、EGFR、Ras、Raf蛋白表达显著降低（P<0.05）；与LRP16-siRNA组比较，外源性EGF组SW620细胞增殖抑制率、E-Cadherin蛋白表达显著降低（P<0.05），克隆形成率、侵袭细胞数、PCNA、N-Cadherin、EGFR、Ras、Raf蛋白表达显著升高（P<0.05）。结论：沉默LRP16表达可能通过抑制EGFR通路活化抑制结肠癌细胞增殖、侵袭及迁移。
- 人白血病相关蛋白16 /
- 结肠癌 /
- 表皮生长因子受体通路 /
- 增殖 /
- 迁移
Abstract: Objective: To study the effect of human leukemia-related protein 16(LRP16) regulating epidermal growth factor receptor(EGFR) pathway on the biological behavior of colon cancer cells. Methods: Colon cancer SW620 cell line was cultured in vitro and divided into negative control group(siRNA-NC group), silent LRP16 expression group(LRP16-siRNA group), exogenous EGF group(LRP16-siRNA+recombinant human EGF group). Real-time fluorescent quantitative PCR(qRT-PCR) method was used to detect the LRP16 mRNA level of SW620 cells in each group; CCK-8 method and plate cloning experiment were used to detect SW620 cell proliferation and clone formation rate in each group; transwell test were used to detect the migration ability of SW620 cells in each group; Western blotting was used to detect the protein expression of proliferating cell nuclear antigen(PCNA), migration and invasion related proteins[E-cadherin, N-cadherin]and pathway related proteins(EGFR, rat sarcoma[Ras], Raf). Results: The cells were successfully transfected with LRP16; compared with the siRNA-NC group, the SW620 cell proliferation inhibition rate and E-Cadherin protein expression in the LRP16-siRNA group were significantly increased(P<0.05), the clone formation rate, scratch healing rate, number of invasive cells, LRP16, PCNA, N-Cadherin, EGFR, Ras, Raf protein expression were significantly reduced(P<0.05); compared with the LRP16-siRNA group, the SW620 cell proliferation inhibition rate and E-Cadherin protein expression in the exogenous EGF group were significantly reduced(P<0.05), the clone formation rate, scratch healing rate, number of invasive cells, PCNA, N-Cadherin, EGFR, Ras, Raf protein expression were significantly increased(P<0.05).Conclusion: Silencing the expression of LRP16 may inhibit the proliferation, invasion and migration of colon cancer cells by inhibiting the activation of the EGFR pathway.
- human leukemia associated protein 16 /
- colon cancer /
- epidermal growth factor receptor pathway /
- proliferation /
- migration

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