摘要
目的研究在不同培养条件下,真核延伸因子2激酶(eEF2K)对三阴性乳腺癌细胞MDA-MB-231增殖的影响,并初步探讨其机制。方法慢病毒感染构建eEF2K表达抑制的MDA-MB-231稳定细胞株以及对应的空载体对照细胞株。在完全、无糖和无血清的培养条件下,SRB法测定细胞增殖曲线,平板克隆实验评价细胞的克隆形成能力,Western blot检测增殖相关蛋白的表达和活性。结果贫营养条件下,抑制MDA-MB-231细胞中eEF2K的表达能够促进细胞生长。而在完全培养条件下,抑制eEF2K表达则抑制细胞的增殖和克隆形成,并降低Akt和mTOR蛋白的活性,对其它增殖相关分子如PI3K、Src、ERK、JAK无明显影响。结论不同培养条件下,eEF2K对MDA-MB-231细胞生长影响不同,其功能值得进一步研究。
Aim To investigate the effects of eukaryotic elongation factor 2 kinase(eEF2K) on the proliferation of triple negative breast cancer cell MDA-MB-231 under different culture conditions and to explore its preliminary mechanism. Methods The lentiviral was used to construct stably transfected cell lines. The cell proliferation curve was determined by SRB assay. The clonogenic capacity of cells was evaluated by plate clone assay. Moreover,to explore its mechanism,expressions and activities of proteins related to cell proliferation were estimated by Western blot assay. Results In complete medium,eEF2K inhibition of MDAMB-231 cells promoted cell proliferation and colony formation,and down-regulated the activities of Akt and m TOR,rather than PI3 K,Src,ERK and JAK. While in nutrient deprivation,inhibiting eEF2K resulted in promoted cell proliferation. Conclusion In different culture conditions,the expression of eEF2K in MDA-MB-231 has different effects on cell growth,and the role of eEF2K in cancer development remains to be explored.
作者
肖敏
王根柱
戚欣
李静
XIAO Min;WANG Gen-zhu;QI Xin;LI Jing(Dept of Molecular Pharmacology,School of Medicine,Ocean University of China,Qingdao Shandong 266000,Chin)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2018年第8期1061-1065,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81673450)