hTERT基因反义核酸促进顺铂诱导原代急性淋巴细胞白血病细胞凋亡的研究  

hTERT gene antisense oligodeoxynucleotide enhances cisplatin-induced apoptosis in ALL cells

作　　者：李文瑜[1] 张洹[2] 何冬梅[2] 

机构地区：[1]广东省人民医院肿瘤中心,广东广州510080 [2]暨南大学医学院血液病研究所,广东广州510632

出　　处：《中国病理生理杂志》2003年第11期1459-1462,共4页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金重点项目 (0 2 1195 ) ;广州市科技计划重点基金 (2 0 0 1-Z - 0 37- 0 1)

摘　　要：目的 :利用人类端粒酶逆转录酶 (hTERT)基因的反义寡核苷酸 (ASPS -ODN)抑制人原代急性淋巴细胞白血病 (ALL)细胞hTERT基因表达后 ,观察顺铂对ALL细胞凋亡的影响。方法 :采用间接免疫荧光标记法通过流式细胞仪检测hTERT蛋白表达量的变化 ;以台盼蓝拒染法计数细胞数 ,确定细胞的增殖情况 ;Hoechst 332 5 8和PI双染色法观察凋亡细胞的形态变化 ;通过流式细胞仪对细胞凋亡峰进行定量分析。结果 :hTERT反义核酸作用于ALL细胞 72hhTERT蛋白表达阳性率为 (32 17± 3 0 0 ) % ,与空白对照组 (6 6 31± 2 84 ) %及正义核酸作用组 (6 2 5 6± 6 11) %比有显著差异 (P <0 0 5 ) ;而正义核酸作用组与空白对照组比无显著差异 (P >0 0 5 )。hTERT反义核酸作用于ALL细胞 2 4h加入顺铂 ,再共同作用 4 8h ,ALL活细胞均数为 1 75 0× 10 8cells/L ,与单用顺铂组 (2 0 90× 10 8cells/L)及hTERT正义核酸联用顺铂组 (1 990× 10 8cells/L)相比 ,对细胞抑制明显增强 (P <0 0 5 )。hTERTASPS -ODN作用于ALL细胞 2 4h再加入顺铂作用 4 8h ,细胞出现典型的凋亡形态学改变。hTERTASPS -ODN与 2 5 μmol/L顺铂联合作用于ALL细胞 4 8h的凋亡细胞百分率 (37 85± 2 4 4 ) %分别同SPS -ODN与顺铂联合作用组 (15 16±2AIM: To explore the effect of human telomerase reverse transcriptase (hTERT) gene antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) on cisplatin-induced apoptosis in cultured primary acute lymphoblastic leukemia (ALL) cells. METHODS: The expression levels of hTERT protein were detected by immunofluorescence using fluoresce isothiocyanate (FITC) lable. Cell surviving fraction was determined using the trypan blue dye exclusion assay. Apoptosis was detected by morphological observation and flow cytomertric cell cycle analysis. RESULTS: The expression of hTERT protein was inhibited after treated by hTERT AS PS-ODN. Treatment with cisplatin after 24 h of exposure to AS PS-ODN had significantly reduced the number of viable ALL cells. However,there was no difference on ALL cells survival between sense oligodeoxynucleotide (S PS-ODN) /CDDP combination and CDDP-treated cells alone. In morphological observation of apoptotic cells using Hoechst 33258 and PI double staining techniques,cells displayed classic apoptotic changes treated with CDDP or CDDP combined with hTERT AS PS-ODN or S PS-ODN at 48 h. Apoptotic rates of cells added CDDP and AS PS-ODN were higher than that of cells added CDDP only ( P <0.05). Apoptotic rates of cells added CDDP and S PS-ODN were similar with that of cells added CDDP only ( P >0.05). CONCLUSION: hTERT AS PS-ODN inhibited the expression of hTERT protein and increased the CDDP-induced apoptosis in primer acute lymphoblastic leukemic cells.

关 键 词：HTERT基因 人类端粒酶逆转录酶基因 反义寡核甘酸类 原代急性淋巴细胞白血病 顺铂 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤]