布比卡因调控LncRNA LBX2-AS1对宫颈癌细胞SiHa增殖及凋亡的影响

Effects of Bupivacaine Regulating the Expression of LncRNA LBX2-AS1 on the Proliferation and Apoptosis of Cervical Cancer Cell SiHa

目的:探讨布比卡因对宫颈癌细胞SiHa增殖、凋亡的影响及其对LncRNA LBX2-AS1的调控作用。方法:体外培养人宫颈癌细胞SiHa,分别加入不同剂量的布比卡因处理细胞,分别将si-NC、si-LBX2-AS1转染至SiHa细胞,分别将pcDNA、pcDNA-LBX2-AS1转染至SiHa细胞后使用布比卡因处理;采用MTT实验检测细胞增殖;应用流式细胞仪检测细胞周期及细胞凋亡率;采用qRT-PCR法检测LBX2-AS1的表达量;Western blot法检测Cleaved-caspase3、pro-caspase3蛋白表达量。结果:布比卡因可明显降低光密度(OD)值、S期细胞比例、LBX2-AS1的表达水平及pro-caspase3蛋白水平(P<0.05),提高凋亡率、G0-G1期细胞比例及Cleaved-caspase3蛋白水平(P<0.05);转染si-LBX2-AS1可明显降低OD值、S期细胞比例及pro-caspase3蛋白水平(P<0.05),提高凋亡率、G0-G1期细胞比例及Cleaved-caspase3蛋白水平(P<0.05);转染pcDNA-LBX2-AS1可明显逆转布比卡因对SiHa细胞增殖、细胞周期及凋亡的作用(P<0.05)。结论:布比卡因可通过下调LBX2-AS1的表达从而抑制宫颈癌细胞增殖、诱导细胞周期阻滞于G0-G1期细胞比例及促进细胞凋亡。

Objective:To explore the effects of bupivacaine on the proliferation and apoptosis of cervical cancer cell SiHa and its regulation to LncRNA LBX2-AS1.Methods:Human cervical cancer cell SiHa was cultured in vitro,and different doses of bupivacaine were added to treat the cells.The si-NC and si-LBX2-AS1 were respectively transfected into SiHa cells.The pcDNA and pcDNA-LBX2-AS1 were transfected into SiHa cells and treated with bupivacaine.MTT assay was used to detect cell proliferation.Flow cytometry was used to detect cell cycle and cell apoptosis rate.The qRT-PCR method was used to detect the expression of LBX2-AS1.Western blot method was used to detect the expressions of Cleaved-caspase3 and pro-caspase3 protein.Results:Bupivacaine could significantly reduce the OD value,the proportion of cells in S phase,the expression level of LBX2-AS1 and the protein level of pro-caspase3(P<0.05),and increase the rate of apoptosis,the proportion of cells in G0-G1 phase and the protein level of Cleaved-caspase3(P<0.05).Transfection of si-LBX2-AS1 could significantly reduce the OD value,the proportion of S-phase cells and the protein level of pro-caspase3(P<0.05),and increase the apoptosis rate,the proportion of cells in G0-G1 phase and the protein level of Cleaved-caspase3(P<0.05).Transfection of pcDNA-LBX2-AS1 could significantly reverse the effects of bupivacaine on the proliferation,cell cycle and apoptosis of SiHa cells(P<0.05).Conclusion:Bupivacaine can inhibit the proliferation of cervical cancer cells,induce cell cycle arrest in the proportion of G0-G1 cells and promote cell apoptosis by down-regulating the expression of LBX2-AS1.