LRP16基因对慢性髓系白血病K562细胞促增殖作用的研究  

Promotive Effect of LRP16 Gene on Proliferation of K562 Cells

作　　者：杨波[1] 卢学春[1] 迟小华[2] 韩为东[3] 于力[4] 楼方定[4] 

机构地区：[1]解放军总医院南楼血液科,北京100853 [2]解放军第262医院药剂科,北京100800 [3]解放军总医院基础医学研究所免疫教研室,北京100853 [4]解放军总医院血液科,北京100853

出　　处：《中国实验血液学杂志》2009年第5期1154-1158,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号30772597);国家自然科学基金资助项目(编号30873086)

摘　　要：本研究探讨超表达LRP16基因对人慢性髓系白血病K562细胞增殖的影响。采用RT-PCR法扩增人LRP16基因开放阅读框(open reading frame,ORF)片段,将其连接到pGEM-T质粒以构建LRP16基因ORF-pGEM-T重组载体。再将测序鉴定过的LRP16基因ORF插入pcDNA3.1+质粒以构建LRP16基因ORF-pcDNA3.1+重组表达质粒,转染K562细胞,建立稳定超表达LRP16基因的K562细胞系。用MTT法测定细胞存活率并绘制生长曲线,流式细胞术检测细胞周期。结果表明:成功扩增出人LRP16基因ORF片段,并构建LRP16基因ORF-pcDNA3.1+重组表达质粒;建立稳定超表达LRP16基因的K562细胞系;超表达LRP16基因具有促进K562细胞增殖的作用,且这种促增殖作用部分是通过促进细胞由G0期进入S期而实现的。结论:超表达LRP16基因可促进K562细胞增殖。The study was aimed to investigate the promotive effect of LRP16 gene on K562 cell proliferation. Open reading frame of LRP16 gene was amplified using reverse transcription-polymerase chain reaction （RT-PCR） and ligated to pGEM-T plasmid to construct LRP16 ORF- pGEM-T recombinant vector. Then, LRP16 ORF identified by sequencing was inserted into pcDNA3.1 ^＋ plasmid to construct LRP16 ORF-pcDNA3.1 ^＋ recombinant expression plasmid which was transfected into K562 cell lines to make overexpression of LRP16 gene in K562 cells. Survival of cells was determined by MTT assay and growth curve of cells was drawn, the cell cycle was detected by flow cytometry. The results showed that LRP16 ORF was successfully amplified, then the LRP16 ORF-pcDNA3.1 ^＋ recombinant plasmid was constructed. The K562 cell line with overexpression of LRP16 gene was established. The promotive effect of LRP16 gene overexpression on proliferation of K562 cells was observed and the effect partially related to the enhancement of cells from GO to S phase induced by LRP16 gene. It is concluded that LRP16 gene overexpression shows a promotive effect on proliferation of K562 cells.

关 键 词：LRP16基因 K562细胞 细胞增殖 慢性髓系白血病 

分 类 号：R733.72[医药卫生—肿瘤]