青藤碱与表柔比星联合处理对黑色素瘤细胞侵袭相关蛋白变化的影响

Effects of Sinomenine combined with Epirubicin on the Changes of Invasion Related Proteins in Melanoma Cells

目的:分析青藤碱(SIN)与表柔比星(EPI)联合处理对黑色素瘤细胞侵袭相关蛋白变化的影响。方法:通过Transwell法对不同条件下黑色素瘤A375细胞侵袭状况进行检测,分为对照组(给予生理盐水)、SIN组(取10μg/m L、100μg/m L分别作为低浓度和高浓度)、EPI组(取5μg/m L、50μg/m L分别作为低浓度和高浓度)、SIN+EPI组(SIN浓度为10μg/m L,EPI浓度为5μg/m L),采用RT-PCR法与Western blot法对各组A375细胞侵袭相关蛋白基质金属蛋白酶-2(MMP-2)及基质金属蛋白酶-9(MMP-9)蛋白及其mRNA的表达进行检测。结果:SIN+EPI组对黑色素瘤A375细胞侵袭的抑制率较SIN组、EPI组均明显升高(P <0.05)。并且,随着SIN、EPI剂量的提高,SIN与EPI对A375细胞侵袭的抑制作用明显提升。相比对照组,SIN组、EPI组、SIN+EPI组MMP-2及MMP-9蛋白及其mRNA水平均受到不同程度的抑制。其中,SIN+EPI组的抑制作用更强(P <0.05)。结论:采用低浓度(10μg/m L)的SIN可有效促进低浓度EPI抑制黑色素瘤细胞侵袭的作用,并且SIN可作为辅助化疗的重要药物,可发挥提升临床疗效的作用。将SIN联合EPI化疗药物,可起到协同、增效的作用,可降低EPI的使用剂量,以防高浓度化疗药物所致的一系列毒副反应,因此具有良好的安全性和有效性。

Objective:To analyze the effect of sinomenine(SIN)combined with epirubicin(EPI)on the changes of invasion-related proteins in melanoma cells.Methods:The invasion status of melanoma A375 cells under different conditions was detected by Transwell.They were divided into control group(given normal saline),SIN group(10 g/mL,100 g/mL as low concentration and high concentration),EPI group(5 g/mL,50 g/mL as low concentration and high concentration),SIN+EPI group(10 g/mL,5 g/mL)and SIN+EPI group(5 g/mL).The expression of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9)proteins and their mRNA were detected by RT-PCR and Western blot.Results:The inhibition rate of SIN+EPI group on the invasion of melanoma A375 cells was significantly higher than that in group SIN and EPI(P<0.05).Moreover,with the increase of SIN and EPI concentration,the inhibition of SIN and EPI on the invasion of A375 cells increased significantly.Compared with the control group,the levels of MMP-2,MMP-9 protein and mRNA in SIN group,the EPI group and the SIN+EPI group were inhibited in varying degrees.Among them,the inhibition effect of SIN+EPI group was higher(P<0.05).Conclusion:The use of low concentration(10μg/mL)SIN can effectively promote the inhibitory effect of low concentration of EPI on the invasion of melanoma cells,and SIN can be used as an important drug for adjuvant chemotherapy.It can play a role in improving the clinical efficacy.SIN combined with EPI chemotherapy drugs can play a synergistic and synergistic effect,and can reduce the dosage of EPI,in order to prevent a series of toxic and side effects caused by high concentration of chemotherapeutic drugs,so it has good safety and effectiveness.