hTERT和β-Catenin在胆囊癌细胞系中的表达及其与癌细胞增殖、侵袭关系研究  被引量:4

Relations of hTERT and β-Catenin Expression with Cell Growth and Invasion in Gallbladder Cell Line

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作  者:丁昂[1] 童赛雄[1] 锁涛[1] 

机构地区:[1]复旦大学附属中山医院普外科,上海200032

出  处:《中国临床医学》2005年第3期452-455,共4页Chinese Journal of Clinical Medicine

摘  要:目的:检测hTERT和β-Catenin在人胆囊癌细胞系中的表达;初步探讨其与癌细胞增殖、侵袭关系。方法:采用端粒酶TRAP、半定量RT-PCR和Western印迹方法检测3种体外培养人胆囊癌细胞系端粒酶活性、hTERT基因和β-Catenin基因mRNA、蛋白质表达水平。四甲基偶氮唑盐(MTT)光吸收法检测琥珀酸脱氢酶(SDH)活性并做细胞增殖计数、运用Transwell小室和划线了解癌细胞侵袭与运动情况。结果:(1)TGBC1TKB、TGDC2TKB和GB-SD端粒酶活性OD值分别为0.183±0.001、0.257±0.002和0.260±0.002;β-Catenm基因mRNA值分别为44718±567、50279±545和50802±371;hTERT基因值分别为51287±1818、62346±733和63345±1154;后两组均高于第1组(P<0.05)。β-Catenm蛋白质值分别为35289±154、32636±345和32510±346;hTERT蛋白质值分别为34515±454、32562±571和32083±163。后两组均低于第1组(P<0.05),有显著差异。(2)TGBC1TKB、TGBC2TKB和GB-SD琥珀酸脱氢酶活性第5天OD值分别为1.324±0.792、1.573±0.(143和1.647±0.033;24h穿膜细胞数分别为60.667±3.512、113.333±5.508和124.667±6.506;24h过线细胞数分别为23.667±1.155、40.000±1.000和42.667±2.082。后两组均高于第1组(P<0.05),有显著差异。结论:端粒酶、hTERT和β-Catenin基因蛋白在人胆囊癌细胞系呈阳性表达;它们可提示胆囊癌细胞的生物学特性,可能影响胆囊癌细胞的增殖、运动和侵袭力。Objective: To explore the relationship of human telomerase reverse transcnptase ( hTERT) and β-Catenin expression with cell growth and invasion in gallbladder carcimoma cell lines, to elucidate their roles in gallbladder carcinogenesis. Methods: (1) Telomerase activity, mRNA and protein of hTERT and β-Catenin in 3 human gallbladder carcinoma cell lines were detected using telomerase TRAP, reverse transcnptase polymerase chain reaction (RT-PCR) and Western blot techniques, respectively. (2) SDH activity by MTT and cell growth, invasion assay by Transwell chamber and migration assay by lineation in culture utensil. Results: (1) The absorbency of the cell (OD value) was used to represent the telomerase activity. In TGBC1TKB, TGBC2TKB and GB-SD, the OD value were 0. 183 ± 0. 001, 0. 257 ± 0. 002 and 0. 260 ± 0. 002, respectively. β-Catenin mRNA value were 44 718 ± 567, 50 279 ± 545 and 50 802 ± 371, respectively. hTERT mRNA value were 51 287 ± 1 818,62 346 ± 733 and 63 345 ± 1 154, respectively. β-Catenin protein value were 35 289 ±154, 32 636 ± 345 and 32 510 ± 346, respectively. hTERT protein value were 34 515 ± 454, 32 562 ± 571 and 32083 ?163, respectively. The OD value, mRNA and protein value of hTERT and β-Catenin in TGBC2TKB and GB-SD than those in TGBC1TKB (P<0. 05). (2) In TG-BC1TKB, TGBC2TKB and GB-SD, the OD value of SDH at 5 days were 1. 324 ± 0.792, 1. 573 ± 0. 043 and 1.647 ± 0. 033, respectively. At 24 h, the counts of cell penetrating through membrane of Transwell chamber were 60. 667 ± 3. 512,113. 333 ± 5. 508 and 124. 667 ± 6. 506, respectively. At 24 h, the counts of cell getting over lineation in culture utensil were 23. 667 ± 1. 155,40. 000 ± 1. (KM) and 42. 667 ± 2.082, respectively. The OD value of SDH, the counts of cell penetrating through membrane of Transwell chamber and getting over lineation in culture utensil were higher in TGBC2TKB and GB-SD than those in TGBC1TKB (P<0. 05). Conclusion: The positive expression of telomerase activity, mRNA and protein of hTERT

关 键 词:胆囊肿瘤 端粒酶逆转录酶 基因 

分 类 号:R735.8[医药卫生—肿瘤]

 

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