乏氧环境对子宫肌瘤细胞增殖及自噬相关蛋白表达的影响

Effects of hypoxia on proliferation and autophagy related protein expression of uterine leiomyoma cells

目的探究乏氧环境对子宫肌瘤细胞增殖和自噬相关蛋白表达的影响。方法选取成都市第三人民医院2017年11月至2018年11月行子宫肌瘤手术治疗的20例病人的子宫肌瘤组织样本(观察组)及周围正常组织(对照组)进行乏氧(1%氧)培养,培养48 h检测两组细胞增殖及自噬相关蛋白的表达差异。结果培养12 h开始至48 h,两组细胞增值率明显下降,并且观察组下降更加显著(P<0.05)。随着缺氧时间增长,两组凋亡细胞比例均显著增加(P<0.05),且观察组细胞凋亡比例更高于对照组(P<0.05)。培养0 h,观察组子宫肌瘤细胞孕激素受体A/B(PR-A/B)(0.436±0.027)/(0.459±0.033)、孕激素受体M(PRM)(0.121±0.013)、缺氧诱导因子1α(HIF-1α)(0.541±0.032)及孔蛋白(porin蛋白)(3.261±0.857)表达水平明显低于对照组PRA/B(0.225±0.022,0.245±0.029)、PR-M(0.084±0.009)、HIF-1α(0.233±0.070)及porin蛋白(0.205±0.027)表达水平(P<0.05)。培养48 h后,观察组PR-A/B、PR-M、HIF-1α及porin蛋白水平显著降低(P<0.05),而对照组各蛋白水平无明显变化(P>0.05)。培养0 h,观察组酵母自噬相关基因(Beclin-1)(0.77±0.08)、选择性自噬接头蛋白(P62)(0.44±0.08)及自噬相关蛋白Ⅱ型微管相关蛋白轻链3(LC3-Ⅱ)(0.55±0.13)显著低于对照组Beclin-1(1.25±0.12)、P62(0.95±0.15)及LC3-Ⅱ(0.81±0.09)表达水平(P<0.05),培养48 h后,观察组各蛋白水平明显升高(P<0.05),而对照组无明显变化(P>0.05)。结论子宫肌瘤细胞增殖及自噬相关蛋白存在明显异常,乏氧环境促进了子宫肌瘤细胞的凋亡和自噬作用。

Objective To explore the effect of hypoxic environment on the proliferation and autophagy related protein expression of uterine leiomyoma cells.Methods The uterine myoma tissue samples(observation group)and normal tissue(control group)of 20 patients who underwent hysteromyoma surgery in Chengdu Third People’s Hospital from November 2017 to November 2018 were selected for hypoxia(1%O_(2))culture.The expression of proliferation and autophagy related protein in the two groups were detected at 48 h.Results From 12 h to 48 h,the cell proliferation rate decreased significantly in the two groups,and the decrease in the observation group was more significant(P<0.05).With the increase of hypoxia time,the proportion of apoptotic cells in both groups increased significantly(P<0.05),and the percentage of apoptotic cells in the observation group was higher than that in the control group(P<0.05).The expression of PR-A/B(0.436±0.027,0.459±0.033),PR-M(0.121±0.013),HIF-1α(0.541±0.032)and porin protein(3.261±0.857)in uterine leiomyoma cells in the observation group was significantly lower than PR-A/B(0.225±0.022,0.245±0.029),PR-M(0.084±0.009),HIF-1α(0.233±0.070)and porin protein(0.205±0.027)in the control group at 0 h(P<0.05).After culture for 48 h,the levels of PR-A/B,PRM,HIF-la and porin protein in the observation group were significantly decreased(P<0.05),but the control group had no significant change in the protein levels(P>0.05).After culture for 0 h,the levels of Beclin-1(0.77±0.08),P62(0.44±0.08)and light chain of microtubule-associated protein typeⅡ(LC3-Ⅱ)(0.55±0.13)in the observation group were significantly lower than Beclin-1(1.25±0.12),P62(0.95±0.15)and LC3-Ⅱ(0.81±0.09)in the control group(P<0.05).After 48 hours of culture,the levels of each protein in the observation group were significantly increased(P<0.05),but there was no significant change in the observation group(P>0.05).Conclusion The proliferation and autophagy related protein of uterine leiomyoma cells are obviously abnormal.Hypoxia environment promotes the apoptosis and autophagy of uterine leiomyoma cells.