Objective Cordycepin is the active component of Cordyceps sinensis and Cordyceps militaris,belongs to Traditional Chinese Medicine,and has a significant anti-cancer activity.This research aimed to study the roles of c...Objective Cordycepin is the active component of Cordyceps sinensis and Cordyceps militaris,belongs to Traditional Chinese Medicine,and has a significant anti-cancer activity.This research aimed to study the roles of cordycepin in inducing apoptosis of cutaneous T-cell lymphoma Hut78 cell line.Methods After treatment with 5 μg/ml cordycepin for 24 hours,the proliferation and apoptosis of Hut78 cells induced by cordycepin were analyzed via MTT and flow cytometry assay,respectively,qRT-PCR and Western blotting were used to investigate the levels of apoptosis-related proteins.Results Proliferation and apoptosis of Hut78 cells were inhibited by cordycepin in a dose-dependent manner.The expression levels of apoptosis-related proteins including caspase-8,cleaved caspase-3,tBid,Bax,Apafl,AIF and DR3 receptor were all decreased in the cells stimulated with 5 μg/ml cordycepin for 24 hours compared with those in the control group.Conclusion Cell proliferation and induced apoptosis could be inhibited by cordycepin via the DR3/mitochondrial signaling pathway.展开更多
文摘目的:探讨人HUT78细胞是否存在褪黑素受体(melatonin receptor,MR)及其亚细胞分布特点。方法:采用异硫氰酸胍-苯酚-氯仿一步法抽提总RNA,通过RT-PCR方法检测MR亚型mt1和MT2的mRNA,并将RT—PCR的阳性产物通过自动测序仪测序;同时应用免疫组化染色方法检测mt1和MT2受体亚型蛋白在HUT78细胞内的分布特点。结果:RT—PCR得到了368 bp
文摘目的探讨合成端粒酶逆转录酶(hTERT)基因及端粒酶RNA(hTR)的小干扰RNA(small interfering RNA,siRNA)对皮肤T细胞淋巴瘤(CTCL)细胞株Hut78端粒酶活性的影响。方法采用体外转录法分别合成两条hTERT—siRNA及hTR—siRNA。采用siRNA直接处理细胞裂解液及磷酸钙共沉淀法将siRNA转染入Hut78细胞两种处理方式,应用端粒酶重复序列扩增聚合酶链反应(TRAP—PCR) 及聚丙烯酰胺凝胶电泳银染检测端粒酶活性的变化。结果体外转录法能高效合成siRNA,产量达 22．4 μg／40μL siRNA合成体系。经250 ng siRNA直接处理细胞裂解液后,可高效降低端粒酶的活性,抑制率达87％左右。30 ng siRNA转染细胞36 h后,可降低端粒酶活性20％左右,而250 ng siRNA可降低 75％左右。结论体外转录法能高效、快速、大量合成siRNA,针对hTERT及hTR基因的siRNA可明显降低Hut78细胞株的端粒酶活性。
文摘Objective Cordycepin is the active component of Cordyceps sinensis and Cordyceps militaris,belongs to Traditional Chinese Medicine,and has a significant anti-cancer activity.This research aimed to study the roles of cordycepin in inducing apoptosis of cutaneous T-cell lymphoma Hut78 cell line.Methods After treatment with 5 μg/ml cordycepin for 24 hours,the proliferation and apoptosis of Hut78 cells induced by cordycepin were analyzed via MTT and flow cytometry assay,respectively,qRT-PCR and Western blotting were used to investigate the levels of apoptosis-related proteins.Results Proliferation and apoptosis of Hut78 cells were inhibited by cordycepin in a dose-dependent manner.The expression levels of apoptosis-related proteins including caspase-8,cleaved caspase-3,tBid,Bax,Apafl,AIF and DR3 receptor were all decreased in the cells stimulated with 5 μg/ml cordycepin for 24 hours compared with those in the control group.Conclusion Cell proliferation and induced apoptosis could be inhibited by cordycepin via the DR3/mitochondrial signaling pathway.
