MARK2 regulates cell polarity and cell proliferation in HeLa cells

Ge Yinggang; Xu Ge; Gao Qing; Liang Xiaoyan

doi:10.13406/j.cnki.cyxb.001360

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MARK2 regulates cell polarity and cell proliferation in HeLa cells

更新时间：2024-01-17

- MARK2 regulates cell polarity and cell proliferation in HeLa cells
- Journal of Chongqing Medical University Vol. 42, Issue 11, Pages: 1483-1487(2017)
- 作者机构：
  
  1. 重庆医科大学附属第一医院胃肠外科
  2. 重庆医科大学生命科学研究院
  3. 重庆医科大学附属第一医院消化内科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001360
  CLC： R730.2
- Published：2017，
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Ge Yinggang, Xu Ge, Gao Qing, et al. MARK2 regulates cell polarity and cell proliferation in HeLa cells. [J]. Journal of Chongqing Medical University 42(11):1483-1487(2017)
DOI：

Ge Yinggang, Xu Ge, Gao Qing, et al. MARK2 regulates cell polarity and cell proliferation in HeLa cells. [J]. Journal of Chongqing Medical University 42(11):1483-1487(2017) DOI： 10.13406/j.cnki.cyxb.001360.

摘要

目的:观察MARK2基因对He La细胞极性与增殖的影响

探讨其可能的机制。方法:脂质体介导MARK2质粒转染He La细胞

G418抗性筛选获得稳定转染的细胞株。TRITC-Phalloidin染色观察细胞极性

平板克隆形成试验研究细胞贴壁生长能力

流式细胞术分析细胞周期各时相细胞的比例

Western blot检测总Rb蛋白及磷酸化Rb蛋白的水平。结果:成功构建稳定转染MARK2的He La细胞系

与空质粒对照组细胞相比

MARK2蛋白表达增强后He La细胞形态改变、上皮细胞极性恢复;MARK2表达增强后He La细胞形成平板克隆数量明显减少[(303.67±7.77)vs.(111.67±7.64)

P=0.000]

细胞周期G1期比例明显增加[(48.82±0.84)%vs.(72.01±2.13)%

P=0.000]、S期比例减少[(36.60±0.58)%vs.(16.03±3.85)%

P=0.010]

Rb蛋白的磷酸化水平明显降低[(7.66±0.74)vs.(1.10±0.13)

t=15.053

P=0.003]。结论:在He La细胞中

MARK2通过抑制Rb蛋白的磷酸化

使细胞周期停滞在G1期

抑制细胞增殖。MARK2能够重建He La细胞上皮极性

协同调控细胞极性与细胞增殖两条通路。

Abstract

Objective:To observe the effect of MARK2 expression on cell polarity and proliferation in He La cells

and to investigate the underlying mechanisms. Methods:A stable He La cell line highly expressing MARK2 protein was established by using G418 selection. Cell polarity was observed with TRITC-Phalloidin staining. Cell proliferation was examined with plate colony formation assay.Proportions of cells in different phases of cell cycle were analyzed by flow cytometry. Levels of total Rb protein and phosphorylated Rb protein were analyzed by Western blot. Results:High expression of MARK2 in Hela stable cells led to phenotypic changes consistent with reversed epithelial polarity. Decreased cell proliferation

increased G1-phase cell proportion

reduced S-phase cell proportion and inhibited Rb phosphorylation were observed in He La stable cells. Conclusion:Enforced expression of MARK2 protein in He La cells leads to decreased Rb phosphorylation

arrests cell cycle at G1-stage thus inhibits cell proliferation. MARK2 can reconstruct epithelial polarity of He La cells

and modulate both signaling pathways regulating cell polarity and cell proliferation.

关键词

细胞周期细胞极性MARK2

Keywords

cell cyclecell polarityMARK2

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