胆汁中端粒酶逆转录酶和Bcl-2基因的表达及意义

目的研究端粒酶逆转录酶(hTERT)和Bcl-2基因在胆管癌患者胆汁中的表达及相互关系;探讨检测胆汁中hTERT和Bcl-2基因对胆管癌的诊断价值。方法将胆总管结石患者胆汁标本20份为对照组,胆管癌患者胆汁标本20份为试验组。采用半定量逆转录-聚合酶链反应(RT-PCR)方法检测标本中hTERT和Bcl-2基因。结果试验组中hTERT基因表达阳性14例,对照组中hTERT基因无表达,2组间差异统计学意义;试验组中Bcl-2基因表达阳性11例,对照组中Bcl-2基因的阳性1例,2组间差异统计学意义;试验组中hTERT和Bcl-2基因表达无相关性。结论胆管癌患者胆汁hTERT基因与Bcl-2基因的表达均明显高于胆总管结石者。端粒酶和Bcl-2在胆管癌胆汁中的表达无相关性。用RT-PCR方法检测胆汁中hTERT和Bcl-2基因的表达对胆管癌的诊断有参考意义。     

Inhibitory effects of antisense phosphorothioate oligodeoxynucleotides on pancreatic cancer cell Bxpc-3 telomerase activity and cell growth in vitro

AIM: To investigate the effect of telomerase hTERT gene antisense oligonucleotide (hTERT-ASO) on proliferation and telomerase activity of pancreatic cancer cell line Bxpc-3. METHODS: MTT assay was used to detect the effect of different doses of hTERT-ASO on proliferation of Bxpc-3 cell for different times. To study the anti-tumor activity, the cells were divided into there groups: Control group (pancreatic cancer cell Bxpc-3); antisense oligonucleotide (hTERT-ASO) group; and nosense oligonucleotide group decorated with phosphorothioate. Telomerase activity was detected using TRAP-PCR-ELISA. Cell DNA distribution was examined using flow cytometry assay. Cell apoptosis was observed by transmission electron microscope in each group. RESULTS: After treatment with 6 mmol/L hTERT-ASO, cell proliferation was inhibited in dose- and time-dependent manner. The telomerase activity decreased after treatment with hTERT-ASO for 72 h. Flow cytometry showed the cell number of G0/G1 phase increased from 2.7% to 14.7%, the cell number of S phase decreased from 72.7% to 51.0%, and a sub-Gl stage cell apoptosis peak appeared in front of G1 stage. CONCLUSION: Telomerase antisense oligodeoxy-nucleotide can inhibit the proliferation of pancreatic cancer cell line Bxpc-3 and decrease the telomerase activity and increase cell apoptosis rate in vitro.     

Caspase-3和hTERT在Barrett’s食管中的表达及其相互关系

目的研究半胱天冬蛋白酶-3（Caspase-3）和端粒酶逆转录酶（hTERT）在Barrett’s食管（Barrett’s esophagus,BE）及食管炎伴贲门腺或胃底腺化生组织中的表达,探讨其在BE发病中的作用。方法应用免疫组织化学法（LDP）检测22例BE和38例食管炎伴贲门腺或胃底腺化生组织中Caspase-3和hTERT的表达,比较两者表达的相关性。结果Caspase-3在BE中的阳性表达率为95.45%,明显高于在食管炎伴贲门腺或胃底腺化生组织中的表达（68.42%）（P=0.000）;hTERT在BE中的阳性表达率为27.27%,明显低于在食管炎伴贲门腺或胃底腺化生组织中的表达（65.79%）（P=0.000）。BE中Caspase-3与hTERT的表达呈强负相关（r=-1,P=0.000）。结论BE中Caspase-3高表达,hTERT低表达。Caspase-3和hTERT在BE发生过程中起重要作用,可能成为预测BE发生的重要参考指标。     

反义 hTERT 基因对 K562 细胞凋亡及周期的影响简

目的：探讨反义hTERT基因抑制K562细胞端粒酶活性对其细胞周期及增殖的影响。方法：用脂质体介导反义hTERT基因和空载体质粒转染K562细胞株,绘制生长曲线,MTT法检测细胞活性,血清饥饿法同步化细胞周期,流式细胞仪检测细胞周期及细胞凋亡率变化。结果：反义hTERT基因转染后,细胞生长明显受到抑制。细胞同步化后,流式细胞仪检测显示：KA组细胞出现早期凋亡峰,细胞阻滞于S期。但正常培养48h,各组细胞无明显凋亡,周期无显著差异。结论：反义hTERT基因可明显抑制K562的增殖,细胞同步化后具有促凋亡作用。