HSV-2型2.2 kb潜伏相关转录子对Hela细胞生长特性的影响

Effect of HSV-2 2.2 kb latency associated transcript on the growth characteristics of Hela cells

摘要:

【目的】研究人单纯疱疹病毒2型(herpes simple virus type 2, HSV-2) 2.2 kb潜伏相关转录体(latency associated transcript, LAT) 对体外培养Hela细胞生长特性的影响。【方法】将真核表达质粒pcDNA3.1-2.2 kb LAT或pcDNA3.1转染Hela细胞后, 观察细胞的生长情况。在4 °C 和43.5 °C应激下, MTT法检测细胞活性, Hoechst 33258染色检测细胞凋亡。【结果】pcDNA3.1-2.2 kb LAT质粒转导入Hela细胞后, 检测到2.2 kb LAT转录; 细胞培养2 d转染pcDNA3.1-2.2 kb LAT细胞明显多于转染pcDNA3.1细胞数, 两者细胞数分别为1.7±0.09×105/mL和1.45±0.14×105/mL, P<0.05。4 °C下 pcDNA3.1-2.2 kb LAT组和pcDNA3.1组细胞凋亡率分别为27.6%±5.7%和7.8%±1.5%, P<0.01; 43.5 °C下pcDNA3.1-2.2 kb LAT组和 pcDNA3.1组细胞凋亡率分别为30.5%±4.6%和5.3%±2.0%, P<0.01。【结论】2.2 kb LAT具有促进细胞增殖, 提高细胞活力和抗凋亡作用, 提示它可能在HSV-2感染中发挥重要作用。

Abstract:

[Objective] In this study, we investigated the influence of HSV-2 2.2 kb latency associated transcript on the growth characteristics of Hela cells in vitro. [Methods] We observed the growth of Hela cells transfected with recombinant expression plasmids pcDNA3.1-2.2 kb LAT along with pcDNA3.1 control. In 4 °C or 43.5 °C, vitality of cells and apoptosis rate of cells was detected by MTT and Hoechst33258 staining. [Results] The transcription of 2.2 kb LAT was detected in Hela cells transfected with pcDNA3.1-2.2 kb LAT. The number of cells transfected with pcDNA3.1-2.2 kb LAT was higher than that transfected with pcDNA3.1 (1.7±0.09×105/mL and 1.45±0.14×105/mL, respectively, P<0.05) at 2 days. The apoptosis rate of cells transfected with pcDNA3.1-2.2 kb LAT was lower than that transfected with pcDNA3.1 (27.6%±5.7%和7.8%±1.5%, respectively, P<0.01) at 4 °C. The apoptosis rate of cells transfected with pcDNA3.1-2.2 kb LAT was lower than that transfected with pcDNA3.1 (30.5%±4.6%和5.3%±2.0%, respectively, P<0.01) at 43.5 °C. [Conclusion] The Hela cells transfected with pcDNA3.1-2.2 kb LAT showed promoted proliferation, heightened vitality and reduced apoptosis. These experiments demonstrated that 2.2 kb LAT could play an important role in HSV-2 infection.