正丁酸钠对肺腺癌细胞增殖、凋亡和Notch信号通路的影响

Effect of sodium butyrate on the proliferation and apoptosis of lung adenocarcinoma cells and Notch signal pathway

目的 探讨正丁酸钠对肺腺癌细胞增殖、凋亡和Notch信号通路的影响。方法 体外培养肺腺癌细胞,采用MTT法检测不同正丁酸钠浓度(0 mmol/L、1.25 mmol/L、2.50 mmol/L、5.00 mmol/L、10.00 mmol/L、20.00 mmol/L和40.00 mmol/L)对肺腺癌细胞生长的抑制效果,筛选出正丁酸钠的有效作用浓度和作用时间进行后续实验,同时设置对照组(不做任何干预)。采用Western blot检测各组肺腺癌细胞的Ki-67、增殖性细胞核抗原(PCNA)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、Notch受体1(NOTCH1)、Hes家族BHLH转录因子1(HES1)、周期素D3(cyclin D3)蛋白表达水平,采用克隆形成实验检测各组肺腺癌细胞的克隆形成率,采用流式细胞术检测各组肺腺癌细胞的凋亡率。结果 随着药物浓度的升高和培养时间的延长,正丁酸钠对肺腺癌细胞的抑制率逐渐升高(均P<0.05),呈现浓度依赖性和时间效应。干预72 h后,对照组、5.00 mmol/L正丁酸钠组、10.00 mmol/L正丁酸钠组、20.00 mmol/L正丁酸钠组的Ki-67、PCNA、Bcl-2、NOTCH1、HES1和cyclin D3蛋白表达水平和克隆形成率均依次降低,Bax蛋白表达水平、细胞凋亡率均依次升高(均P<0.05)。结论 正丁酸钠可抑制肺腺癌细胞的生长、增殖并促进其凋亡,这可能与其抑制增殖相关蛋白和Notch信号通路相关蛋白的表达,以及促进凋亡相关蛋白的表达有关。

Objective To explore the effect of sodium butyrate on the proliferation and apoptosis of lung adenocarcinoma cells and Notch signaling pathway.Methods The lung adenocarcinoma cells were cultured in vitro.The inhibitory effect of different concentrations of sodium butyrate(0 mmol/L,1.25 mmol/L,2.50 mmol/L,5.00 mmol/L,10.00 mmol/L,20.00 mmol/L,and 40.00 mmol/L) on the growth of lung adenocarcinoma cells was detected by the MTT method to screen the effective concentrations and time of sodium butyrate for the following experiment;simultaneously,the control group was set up(without any intervention).The expressions of Ki-67,proliferating cell nuclear antigen(PCNA),B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),Notch receptor 1(NOTCH1),Hes family BHLH transcription factor 1(HES1) and cyclin D3 proteins were detected by the Western blot.The colony formation rate of lung adenocarcinoma cells was detected by the colony formation assay.The apoptosis rate of lung adenocarcinoma cells was detected by the flow cytometry.Results With the increase of drug concentration and extension of culture time,the inhibition rate of sodium butyrate on lung adenocarcinoma cells was increased(all P<0.05),and it interpreted concentration dependence and time effect.The expressions of Ki-67,PCNA,Bcl-2,NOTCH1,HES1 and cyclin D3 proteins,as well as the colony formation rate were decreased successively,while the expressions of Bax protein,and cell apoptosis rate were elevated successively in the control group,sodium butyrate of 5.00 mmol/L group,sodium butyrate of 10.00 mmol/L group,and sodium butyrate of 20.00 mmol/L group after 72-hour intervention(all P<0.05).Conclusion Sodium butyrate can inhibit the growth and proliferation of lung adenocarcinoma cells,and promote their apoptosis,which may be related to inhibiting the expressions of proliferation-related proteins and Notch signaling pathway related proteins,and to promoting expressions of apoptosis related proteins.