高糖状态下甲状旁腺素受体1对乳腺癌的作用机制研究

Effect of parathyroid hormone 1 receptor on SHZ-88 cell under high glucose condition

目的阐明高糖状态下甲状旁腺受体1(PTH1R)对乳腺癌细胞株SHZ-88的作用机制。方法应用实时(real time)PCR分别检测0(对照组)、5、15、25mmol/L葡萄糖浓度下PTH1R mRNA水平;构建出PTH1R基因沉默(siPTH1R)的细胞模型后,分别应用MTT法、TUNEL-FITC/Hoechst33258及Western blot法检测对照组、25mmol/L葡萄糖处理组(高糖组)、25mmol/L葡萄糖处理的阴性PTH1R基因序列组(高糖siPTH1R-NC组)及高糖阳性PTH1R基因序列组(高糖siPTH1R组)细胞活力、细胞凋亡情况及Bax、Bcl-2蛋白的表达。结果随着葡萄糖浓度升高,PTH1R mRNA水平增高,其中25mmol/L葡萄糖处理后PTH1RmRNA水平最高(均P<0.01);高糖siPTH1R组细胞活力(P<0.05,P<0.01及P<0.01)及Bcl-2表达(均P<0.01)低于对照组、高糖组及高糖siPTH1R-NC组;高糖siPTH1R组中细胞凋亡水平及Bax表达高于对照组、高糖组及高糖siPTH1R-NC组(均P<0.01);与对照组比较,高糖组细胞活力(P<0.01)及Bcl-2表达(P<0.01)增高,而Bax表达(P<0.01)降低。结论高糖状态下PTH1R表达水平与SHZ-88细胞增殖能力有关,抑制PTH1R表达可能为糖尿病合并乳腺癌治疗有效靶点之一。

Objective To clarify the effect of PTH1 Ron SHZ-88 cell under high glucose.Methods The expression of PTH1 R mRNA of 0,5,15 and 25mmol/L glucose groups were detected by realtime PCR,After the successful construction of recombinant PTH1R-siRNA vectors established in SHZ-88 cell model.Cell viability,cell apoptosis and the expression of Bax and Bcl-2of Control groups（0mmol/L glucose）,high glucose（25mmol/L glucose）,high glucose＋siPTH1R-Negative control group and high glucose＋siPTH1Rgroups were measured by MTT,AnnexinⅤ and Western blot in SHZ-88 cell.Results SHZ-88 cell by stimulated 25mmol/L glucose significantly promoted the level of PTH1RmRNA（all P〈0.01）,this promotion effect is concentration dependent.Compared with Control groups,high glucose groups and high glucose＋siPTH1R-NC control groups,the cell viability and the expression of Bcl-2decreased in high glucose＋siPTH1Rgroups（P〈0.05,P〈0.01 and P〈0.01）,but cell apoptoisis and the expression of Bax increased（all P〈0.01）.High glucose groups had lower cell apoptosis and higher cell viability（P〈0.01）level than others.High glucose could up-regulate the expression of Bcl-2and down-regulate the expression of Bax（all P〈0.01）.Conclusion There is a closely relationship between PTH1 Rand cell proliferation which is under the condition of high glucose,PTH1 R maybe a good therapy target of diabetes complicat with breast cancer.