两歧双歧杆菌胞外多糖对胃癌细胞及端粒酶逆转录酶的影响

[目的]研究从双歧杆菌属两歧双歧杆菌(Bifidobacterium bifidum)提取的细胞表面成分胞外多糖(Exopolysaccharide, B. EPS)对人胃癌细胞BGC-823的生长抑制作用及对端粒酶限速因子hTERT活性的影响.[方法]将3种不同浓度B.EPS体外作用于胃癌细胞BGC-823,MTT法检测细胞生长抑制率并辅以形态学观察;异硫氰酸盐(FITC)联合PI染色,通过流式细胞术检测肿瘤细胞初期调亡情况;肿瘤细胞端粒酶限速因子hTERT mRNA经RT-PCR检测B.EPS对端粒酶活性抑制作用;通过荧光分光光度计显示B.EPS对胃癌细胞作用后胞内Ca2+含量变化.[结果]经过检测发现,B.EPS对人胃癌细胞BGC823的生长显著抑制(P<0.05)呈剂量时间反应关系;细胞中hTERT mRNA在B.EPS的作用下表达降低(P<0.05),有一定剂量效应关系;随着B.EPS对肿瘤细胞的抑制,细胞内Ca2+含量显著增加(P<0.05).[结论] B.EPS诱导人胃癌细胞BGC-823调亡的机制可能与改变肿瘤细胞端粒酶限速因子hTERT mRNA表达量和细胞内钙离子浓度有关.     

Variant forms of upstream stimulatory factors (USFs) control the promoter activity of hTERT,the human gene encoding the catalytic subunit of telomerase

It is known that Myc regulates the expression of TERT, the telomerase catalytic subunit gene, by binding to E box. Here we show that another E box-binding protein, upstream stimulatory factor (USF), also regulates TERT expression. Specifically, the N-terminally truncated form of USF2 is present in telomerase-negative/resting human lymphocytes, but not in telomerase-positive/phytohemagglutinin-activated lymphocytes. In electrophoretic mobility shift assay, both full-length and truncated USF2s bound to the TERT E box. In a transient expression assay, the truncated USF had a dominant-negative effect on both exogenous full-length USF and endogenous positive regulators for activating TERT expression. These results suggest that the differential abundance of truncated USF2 may regulate telomerase activity during lymphocyte activation.