鼠尾草酚通过PPARγ信号通路调节膀胱癌T24细胞增殖及侵袭机制研究

Study on Mechanism of Carnosol Regulating Proliferation and Invasion of Bladder Cancer T24 Cells by PPARγ Signal Pathway

目的:探讨鼠尾草酚调节膀胱癌T24细胞增殖及侵袭的分子机制。方法:将膀胱癌T24细胞分为T24细胞组、5-氟尿嘧啶组及鼠尾草酚低、高剂量组(低、高剂量组)。5-氟尿嘧啶组加入50.0μg/mL的5-氟尿嘧啶,低、高剂量组分别加入50.0μg/mL、100.0μg/mL的鼠尾草酚,培养72 h。测定各组细胞增殖情况、克隆形成数目、侵袭迁移水平、凋亡水平及过氧化物酶体增殖物激活受体γ(PPARγ)、β-连环蛋白(β-catenin) mRNA与蛋白水平。结果:与T24细胞组比较,其余各组细胞凋亡率、PPARγm RNA及蛋白表达水平升高(P<0.05),OD值、存活率、细胞克隆形成数目、穿膜数、β-catenin mRNA及蛋白表达水平降低(P<0.05)。与5-氟尿嘧啶组比较,低剂量组细胞凋亡率、PPARγmRNA及蛋白表达水平降低(P<0.05),OD值、存活率、细胞克隆形成数目、穿膜数、β-catenin mRNA及蛋白表达水平升高(P<0.05);高剂量组细胞凋亡率、PPARγmRNA及蛋白表达水平升高(P<0.05),OD值、存活率、细胞克隆形成数目、穿膜数、β-catenin m RNA及蛋白表达水平降低(P<0.05)。与低剂量组比较,鼠尾草酚高剂量组细胞凋亡率、PPARγmRNA及蛋白表达水平升高(P<0.05),OD值、存活率、细胞克隆形成数目、穿膜数、β-catenin mRNA及蛋白表达水平降低(P<0.05)。结论:鼠尾草酚能抑制膀胱癌T24细胞增殖、侵袭迁移,促进膀胱癌T24细胞凋亡;其机制可能与鼠尾草酚能促进PPARγmRNA及蛋白表达,激活PPARγ信号通路进而抑制β-catenin mRNA和蛋白表达有关。

Objective: To investigate the molecular mechanism of carnosol regulating the proliferation and invasion of bladder cancer T24 cells. Methods:Bladder cancer T24 cells were respectively divided into the T24 cell group,the5-fluorouracil group and the carnosol groups with low and high dose(the low-dose group and the high-dose group).The 5-fluorouracil group was given 50.0 μg/mL of 5-fluorouracil,and the low-dose and high-dose groups were given 50.0 μg/mL and 100.0 μg/mL of carnosol respectively. All groups were cultured for 72 hours. Cell proliferation,the number of colony formation as well as the levels of invasion and migration, apoptosis, peroxisome proliferator activated receptor γ(PPARγ),β-catenin m RNA and protein in each group were measured. Results:Compared with those in the T24 cell group,the apoptosis rates and the levels of PPARγ m RNA and protein expressions in the other groups were increased(P<0.05);the OD value,survival rate,the number of cell clone formation and cell membrane penetration,and the levels ofβ-catenin mRNA and protein expressions were decreased(P<0.05). Compared with those in the 5-fluorouracil group,the apoptosis rate and the levels of PPARγ mRNA and protein expressions in the low-dose group were decreased(P<0.05),and the OD value,survival rate,the numbers of cell clone formation and cell membrane penetration,and the levels of β-catenin mRNA and protein expressions were increased(P<0.05);the apoptosis rate and the levels of PPARγ mRNA and protein expressions in the high-dose group were increased(P<0.05),and the OD value,survival rate,the number of cell clone formation and cell membrane penetration,and the levels of β-catenin mRNA and protein expressions were decreased(P<0.05). Compared with those in the low-dose group, the apoptosis rate and the levels of PPARγ mRNA and protein expressions in the high-dose group were increased(P<0.05),and the OD value,survival rate,the number of cell clone formation and cell membrane penetration,and the levels of β-catenin mRNA and protein expressions were decreased(P<0.05). Conclusion:Carnosol can inhibit proliferation,invasion and migration of bladder cancer T24 cells and promote their apoptosis. Its mechanism may be related to the promotion of PPAR mRNA and protein expressions, activation of PPARγsignal pathway by carnosol,and therefore inhibit β-catenin mRNA and protein expressions.