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目的研究Cr(Ⅵ)对果蝇S2细胞凋亡的影响,初步探讨其可能的凋亡机制。方法将果蝇S2细胞分别暴露于含Cr(Ⅵ)终浓度为0(对照)、50、100、200、400、800μmol/L的新鲜培养基培养12 h。检测果蝇S2细胞的凋亡情况和凋亡相关基因Debcl、Buffy、Drice、P53 mRNA的表达水平及Drice的相对活化程度。结果 Cr(Ⅵ)处理浓度为400μmol/L时可观察到S2细胞染色质浓染、出现凋亡小体,当Cr(Ⅵ)浓度达到800μmol/L时,可明显观察到细胞崩解及坏死;与对照组比较,各剂量Cr(Ⅵ)染毒组果蝇S2细胞的早期凋亡率均较高,差异有统计学意义(P<0.05)。当Cr(Ⅵ)浓度为50~400μmol/L时,果蝇S2细胞的早期凋亡率随着Cr(Ⅵ)浓度的上升而升高;当Cr(Ⅵ)浓度达到800μmol/L时,果蝇S2细胞的早期凋亡率下降。与对照组比较,400μmol/L Cr(Ⅵ)染毒组果蝇S2细胞Debcl、Drice mRNA的表达水平均较低,P53 mRNA的表达水平均较高,差异有统计学意义(P<0.05);而Buffy mRNA的表达水平无明显改变,Drice相对活化程度降低(P<0.01)。结论天冬氨酸蛋白水解酶(Caspases)并未参与高浓度下Cr(Ⅵ)诱导果蝇S2细胞的凋亡过程,其凋亡机制可能与P53基因相关。
Aim To investigate the effect of Cr (Ⅵ) on apoptosis of Drosophila S2 cells and to explore its possible mechanism of apoptosis. Methods Drosophila S2 cells were exposed to Cr (Ⅵ) final concentration of 0 (control), 50,100,200,400,800μmol / L of fresh medium for 12 h. The apoptosis of Drosophila S2 cells and the expressions of Debcl, Buffy, Drice and P53 mRNA and the relative activation of Drice were detected. Results Chromosomal staining of S2 cells was observed when the concentration of Cr (Ⅵ) was 400 μmol / L. Apoptotic bodies were observed. When Cr (Ⅵ) concentration reached 800 μmol / L, cell disintegration and necrosis were observed obviously. Compared with the control group, the early apoptotic rate of S2 cells in Cr (Ⅵ) -treated Drosophila was higher than that in the control group, with significant difference (P <0.05). When Cr (Ⅵ) concentration was 50 ~ 400μmol / L, the early apoptotic rate of Drosophila S2 cells increased with the increase of Cr (Ⅵ) concentration. When the concentration of Cr (Ⅵ) reached 800μmol / L, The early apoptosis rate of S2 cells decreased. Compared with the control group, the expressions of Debcl and Drice mRNA in S2 cells treated with 400μmol / L Cr (Ⅵ) were lower and the expression of P53 mRNA were higher, the difference was statistically significant (P <0.05). However, the expression level of Buffy mRNA did not change significantly and the relative activation of Drice decreased (P <0.01). Conclusion Caspases were not involved in Cr (Ⅵ) -induced apoptosis in Drosophila S2 cells at high concentrations, and their apoptosis mechanisms may be related to P53 gene.