参一胶囊抑制人非小细胞肺癌PC14细胞增殖的信号通路作用机制研究

Mechanism analysis of signaling pathway of Shenyi capsule inhibiting proliferation of human non-small cell lung cancer PC14 cells

目的:分析参一胶囊抑制人非小细胞肺癌PC14细胞增殖的信号通路作用机制。方法:采用浓度分别为0 mmol/L、4 mmol/L、8 mmol/L、16 mmol/L的参一胶囊对PC14细胞进行24 h、48 h和72 h的处理,采用流式细胞术和MTT法检测细胞的增殖和凋亡情况。采用5μmol/L的SP600125(JNK抑制剂)与SB203580(ERK抑制剂)对PC14细胞进行30 min处理,并加入不同浓度的参一胶囊继续培养24 h、48 h和72 h培养,观察细胞增殖情况,并检测参一胶囊对JNK和ERK活化的影响。结果:与0 mmol/L浓度组(对照组)比较,PC14细胞采用不同浓度的参一胶囊处理24 h、48 h和72 h后,4 mmol/L组、8 mmol/L组、16 mmol/L组的细胞活力均降低,且随着参一胶囊浓度的增大,细胞活力逐渐降低。浓度为16 mmol/L时,细胞活力最低(P<0.05)。与0 mmol/L浓度组比较,4 mmol/L组、8 mmol/L组、16 mmol/L组的细胞总凋亡率分别为(2.99±1.25)%、(15.37±4.95)%、(19.63±6.00)%,且随着参一胶囊浓度的增大,细胞凋亡率逐渐升高。浓度为16 mmol/L时,细胞凋亡率最高(P<0.05)。与0 mmol/L浓度组比较,处理48 h后,4 mmol/L组、8 mmol/L组、16 mmol/L组PC14细胞处于各期的比例无明显差异(P>0.05)。SP600125与SB203580均能有效抑制参一胶囊诱导的细胞凋亡,单纯加入参一胶囊与加入SP600125或SB203580的组别进行比较,细胞活力均较低(P<0.05)。PC14细胞经过不同浓度的参一胶囊处理后,p-p38MAPK蛋白的表达并未受到药物的影响,但随着药物浓度的增加p-JNK和p-ERK水平呈现降低趋势。结论:参一胶囊通过诱导JNK和ERK表达能够促进其磷酸化,且随着浓度的增大,可导致细胞活力明显降低,细胞凋亡率逐渐升高,p-JNK和p-ERK水平逐渐降低,对非小细胞肺癌PC14细胞增殖有明显的抑制作用。

Objective:To analyze the mechanism of signal pathway of Shenyi Capsule in inhibiting the proliferation of human non-small cell lung cancer PC14 cells.Methods:PC14 cells were treated with Shenyi capsules at concentrations of 0 mmol/L,4 mmol/L,8 mmol/L,and 16 mmol/L for 24 h,48 h,and 72 h.Flow cytometry and MTT were used to detect cell proliferation and apoptosis,respectively.Then use 5μmol/L SP600125(JNK inhibitor)and SB203580(ERK inhibitor)to treat PC14 cells for 30 min,and add different concentrations of Shenyi capsules to continue culturing for 24 h,48 h and 72 h,observe the cell proliferation,and detect the effect of Shenyi capsule on JNK and ERK activation.Results:Compared with the 0 mmol/L group(control group),the cell viability of 4 mmol/L group,8 mmol/L group,and 16 mmol/L group was decreased,and with the increase of the concentration of Zhushenyi capsule,the cell viability gradually decreased.When the concentration was 16 mmol/L,the cell viability was the lowest(P<0.05).Compared with the control group,the total apoptosis rates of 4 mmol/L group,8 mmol/L group and 16 mmol/L group were(2.99±1.25)%,(15.37±4.95)%,and(19.63±6.00)%.As the concentration of Shenyi capsule increased,the cell apoptosis rate gradually increased.When the concentration was 16 mmol/L,the cell apoptosis rate was the highest(P<0.05).Compared with the control group group,after 48 hours of treatment,the proportions of PC14 cells in the 4 mmol/L group,8 mmol/L group,and 16 mmol/L group showed no significant difference.Both SP600125 and SB203580 could effectively inhibit the cell apoptosis induced by Shenyi capsule.Compared with the group that added Shenyi capsule alone and group that added SP600125 or SB203580,the cell viability was low(P<0.05).It was found that the expression of p-p38MAPK protein was not affected by the drug after PC14 cells were treated with different concentrations of Shenyi capsules,but with the increase of drug concentration,p-JNK and the level of p-ERK showed decreasing trend.Conclusion:Shenyi capsules can promote the phosphorylation by inducing the expression of JNK and ERK,and with the increase of the concentration,the cell activity significantly reduces,the apoptosis rate gradually increases,and the p-JNK and P-ERK levels gradually decreases,which has an obvious inhibitory effect on the proliferation of PC14 cells in non-small cell lung cancer.