目的 :探讨Bmi1如何调控舌鳞癌侧群细胞的迁移、侵袭和增殖能力。方法 :首先采用Hoechst33342法,利用流式细胞仪分选舌鳞癌细胞株UM1(高转移株)中的侧群细胞(side population cell,SP)和非侧群细胞(non-side population cell,Non-SP)。T...目的 :探讨Bmi1如何调控舌鳞癌侧群细胞的迁移、侵袭和增殖能力。方法 :首先采用Hoechst33342法,利用流式细胞仪分选舌鳞癌细胞株UM1(高转移株)中的侧群细胞(side population cell,SP)和非侧群细胞(non-side population cell,Non-SP)。Transwell实验检测沉默Bmi1后SP细胞的迁移、侵袭能力。球囊形成和平板克隆实验检测沉默Bmi1后SP细胞的球囊和克隆形成率;CCK8实验检测沉默Bmi1后SP细胞的增殖能力。Western免疫印迹检测沉默Bmi1后SP细胞中侵袭、转移相关基因(SOD2、Slug)及干细胞标志物(ABCG2、Nanog)的表达水平。采用SPSS17.0软件包对数据进行统计学处理。结果:转染Bmi1 si RNA使SP细胞中Bmi1表达水平下调后,SP细胞的迁移和侵袭能力显著下降;球囊形成率和克隆形成率也显著低于对照组;增殖能力受到抑制;SOD2、Slug和干细胞标志物ABCG2和Nanog的表达水平显著下降。结论 :沉默Bmi1可调控舌鳞癌侧群细胞的迁移、侵袭和增殖。展开更多
The intestinal epithelial lining plays a central role in the digestion and absorption of nutrients,but exists in a harsh luminal environment that necessitates continual renewal.This renewal process involves epithelial...The intestinal epithelial lining plays a central role in the digestion and absorption of nutrients,but exists in a harsh luminal environment that necessitates continual renewal.This renewal process involves epithelial cell proliferation in the crypt base and later cell migration from the crypt base to the luminal surface.This process is dependent on multi-potent progenitor cells,or stem cells,located in each crypt.There are about 4 to 6 stem cells per crypt,and these stem cells are believed to generate distinct end-differentiated epithelial cell types,including absorptive cells,goblet cells,enteroendocrine cells and Paneth cells,while also maintaining their own progenitor cell state.Earlier studies suggested that intestinal stem cells were located either in the crypt base interspersed between the Paneth cells [i.e.crypt base columnar(CBC) cell model] or at an average position of 4 cells from the crypt base [i.e.label-retaining cells(LRC +4) model].Recent studies have employed biomarkers in the in vivo mammalian state to more precisely evaluate the location of these progenitor cells in the intestinal crypt.Most notable of these novel markers are Lgr5,a gene that encodes a G-protein-coupled receptor with expression restricted to CBC cells,and Bmi 1,which encodes a chromatin remodeling protein expressed by LRC.These studies raise the possibility that there may be separate stem cell lines or different states of stem cell activation involved in the renewal of normal mammalian intestinal tract.展开更多
Objective： The aim of the study was to observe the effect of Bmil reduction on the self-renewal and tumorigenicity ability of lung cancer stem cells （LCSCs） in human lung adenocarcinoma. Methods： Human lung adenoc...Objective： The aim of the study was to observe the effect of Bmil reduction on the self-renewal and tumorigenicity ability of lung cancer stem cells （LCSCs） in human lung adenocarcinoma. Methods： Human lung adenocarcinoma cells A549 were consecutively passaged in NOD/SCID mice treated with Paclitaxel weekly. The proportions of LCSCs in A549 cells and the cells from the third passage （A549-3rd） were compared. The expression of Bmil in LCSCs was silenced by intratumoral injection with lentivirus-delivered Broil small hairpin RNA （shRNA）. RT-PCR and Western blot were used to test the mRNA and protein expressions of Broil in LCSCs. The protein level of p16INK4A was analyzed by Western blotting. The self- renewal and tumorigenicity ability of LCSCs were evaluated by counting the sphere formation rate in serum-free medium and the tumor formation rate in NOD/SCID mice. Results： In vivo passaging ofA549 cells under chemotherapy pressure enriched for LCSCs. The expression of Broil in LCSCs increased. Down-regulation of Bmil by RNA interference resulted in reduced self-renewal and tumorigenicity ability of LCSCs and paralleled the increased expression of p16INK4A, a Bmil target. Conclu- sion： Broil regulates self-renewal and tumorigenicity of LCSCs by silencing some target genes, including p16INK4A.展开更多
文摘目的 :探讨Bmi1如何调控舌鳞癌侧群细胞的迁移、侵袭和增殖能力。方法 :首先采用Hoechst33342法,利用流式细胞仪分选舌鳞癌细胞株UM1(高转移株)中的侧群细胞(side population cell,SP)和非侧群细胞(non-side population cell,Non-SP)。Transwell实验检测沉默Bmi1后SP细胞的迁移、侵袭能力。球囊形成和平板克隆实验检测沉默Bmi1后SP细胞的球囊和克隆形成率;CCK8实验检测沉默Bmi1后SP细胞的增殖能力。Western免疫印迹检测沉默Bmi1后SP细胞中侵袭、转移相关基因(SOD2、Slug)及干细胞标志物(ABCG2、Nanog)的表达水平。采用SPSS17.0软件包对数据进行统计学处理。结果:转染Bmi1 si RNA使SP细胞中Bmi1表达水平下调后,SP细胞的迁移和侵袭能力显著下降;球囊形成率和克隆形成率也显著低于对照组;增殖能力受到抑制;SOD2、Slug和干细胞标志物ABCG2和Nanog的表达水平显著下降。结论 :沉默Bmi1可调控舌鳞癌侧群细胞的迁移、侵袭和增殖。
文摘The intestinal epithelial lining plays a central role in the digestion and absorption of nutrients,but exists in a harsh luminal environment that necessitates continual renewal.This renewal process involves epithelial cell proliferation in the crypt base and later cell migration from the crypt base to the luminal surface.This process is dependent on multi-potent progenitor cells,or stem cells,located in each crypt.There are about 4 to 6 stem cells per crypt,and these stem cells are believed to generate distinct end-differentiated epithelial cell types,including absorptive cells,goblet cells,enteroendocrine cells and Paneth cells,while also maintaining their own progenitor cell state.Earlier studies suggested that intestinal stem cells were located either in the crypt base interspersed between the Paneth cells [i.e.crypt base columnar(CBC) cell model] or at an average position of 4 cells from the crypt base [i.e.label-retaining cells(LRC +4) model].Recent studies have employed biomarkers in the in vivo mammalian state to more precisely evaluate the location of these progenitor cells in the intestinal crypt.Most notable of these novel markers are Lgr5,a gene that encodes a G-protein-coupled receptor with expression restricted to CBC cells,and Bmi 1,which encodes a chromatin remodeling protein expressed by LRC.These studies raise the possibility that there may be separate stem cell lines or different states of stem cell activation involved in the renewal of normal mammalian intestinal tract.
基金Supported by grants from the National Natural Science Foundation of China (No. 30772144)Natural Science Foundation of Chongqing (No. CSTC, 2009BB5148)
文摘Objective： The aim of the study was to observe the effect of Bmil reduction on the self-renewal and tumorigenicity ability of lung cancer stem cells （LCSCs） in human lung adenocarcinoma. Methods： Human lung adenocarcinoma cells A549 were consecutively passaged in NOD/SCID mice treated with Paclitaxel weekly. The proportions of LCSCs in A549 cells and the cells from the third passage （A549-3rd） were compared. The expression of Bmil in LCSCs was silenced by intratumoral injection with lentivirus-delivered Broil small hairpin RNA （shRNA）. RT-PCR and Western blot were used to test the mRNA and protein expressions of Broil in LCSCs. The protein level of p16INK4A was analyzed by Western blotting. The self- renewal and tumorigenicity ability of LCSCs were evaluated by counting the sphere formation rate in serum-free medium and the tumor formation rate in NOD/SCID mice. Results： In vivo passaging ofA549 cells under chemotherapy pressure enriched for LCSCs. The expression of Broil in LCSCs increased. Down-regulation of Bmil by RNA interference resulted in reduced self-renewal and tumorigenicity ability of LCSCs and paralleled the increased expression of p16INK4A, a Bmil target. Conclu- sion： Broil regulates self-renewal and tumorigenicity of LCSCs by silencing some target genes, including p16INK4A.
