藻蓝蛋白对非小细胞肺癌细胞移植瘤生长的抑制及促凋亡作用的机制研究

Study on the Mechanism of Phycocyanin on the Growth of Non-small Cell Lung Cancer Cell Xenografts and Its Pro-apoptotic Effect

目的:探讨藻蓝蛋白对非小细胞肺癌细胞移植瘤生长的抑制及促凋亡作用的机制研究。方法:4~6周龄30只无胸腺BALB/c裸鼠随机分为移植瘤组和藻蓝蛋白组。每5天用卡尺测量裸鼠肿瘤体积。通过RT-PCR检测裸鼠肿瘤中凋亡相关因子MMP-2、MMP-9、Bcl-2、Bax的m RNA表达。通过流式细胞术分析细胞周期。通过蛋白印迹分析EMT相关蛋白的表达。通过ELISA检测血浆细胞因子TNF-α、IL-6、IL-1β和TGF-β的浓度。通过蛋白印迹分析STAT3/NF-κB信号通路的表达。结果:第13天时,移植瘤组和蓝藻蛋白组肿瘤大小比较无差异(P>0.05),第18天和第25天时,藻蓝蛋白组肿瘤体积较移植瘤组减小(P<0.05)。藻蓝蛋白组MMP-2、MMP-9和Bcl-2的m RNA表达较移植瘤组降低(P<0.05),藻蓝蛋白组Bax m RNA表达较移植瘤组升高(P<0.05)。藻蓝蛋白组G1期占比较移植瘤组升高(P<0.05),藻蓝蛋白组G1期占比较移植瘤组升高(P<0.05),藻蓝蛋白组S期和G2期占比较移植瘤组降低(P<0.05)。藻蓝蛋白组N-钙粘蛋白和VEGF蛋白表达较移植瘤组降低(P<0.05),藻蓝蛋白组E-钙粘蛋白表达较移植瘤组升高(P<0.05)。藻蓝蛋白组TNF-α、IL-6、IL-1β和TGF-β的浓度较移植瘤组降低(P<0.05)。藻蓝蛋白组p-STAT3、p-IκBα和p-NF-κB p65蛋白表达较移植瘤组降低(P<0.05)。结论:藻蓝蛋白通过调控STAT3/NF-κB信号通路抑制炎性细胞因子的分泌和EMT发生,促进肿瘤细胞凋亡,抑制裸鼠体内移植瘤的生长。

Objective:To investigate the mechanism of phycocyanin’s inhibitory effect on the growth of non-small cell lung cancer cell transplanted tumors and the mechanism of promoting apoptosis.Methods:Thirty 4-6 weeks old athymic BALB/c nude mice were randomly divided into transplanted tumor group and phycocyanin group.The tumor volume of nude mice was measured with a caliper every 5 days.The m RNA expression of apoptosis-related factors MMP-2,MMP-9,Bcl-2 and Bax in nude mouse tumors was detected by RT-PCR.The cell cycle was analyzed by flow cytometry.The expression of EMT-related proteins was analyzed by Western blot.The plasma cytokines TNF-α,IL-6,IL-1βand TGF-βconcentrations were detected by ELISA.The expression of STAT3/NF-κB signaling pathway was analyzed by Western blot.Results:On the 13th day,there was no difference in tumor size between the transplanted tumor group and the cyanobacterial protein group(P>0.05).On the 18th and 25th days,the tumor volume of the phycocyanin group was smaller than that of the transplanted tumor group(P<0.05).The m RNA expression of MMP-2,MMP-9 and Bcl-2 in the phycocyanin group was lower than that in the transplanted tumor group(P<0.05),and the expression of Bax m RNA in the phycocyanin group was higher than that in the transplanted tumor group(P<0.05).Phycocyanin group G1 stage accounted for a higher rate than transplanted tumor group(P<0.05),phycocyanin group G1 stage accounted for a higher rate than transplanted tumor group(P<0.05),phycocyanin group S stage and G2stage accounted for more transplantation The tumor group decreased(P<0.05).The expression of N-cadherin and VEGF protein in the phycocyanin group was lower than that in the transplanted tumor group(P<0.05),and the expression of E-cadherin in the phycocyanin group was higher than that in the transplanted tumor group(P<0.05).The concentrations of TNF-α,IL-6,IL-1βand TGF-βin the phycocyanin group were lower than those in the transplanted tumor group(P<0.05).The expression of p-STAT3,p-IκBαand p-NF-κB p65 protein in the phycocyanin group was lower than that in the xenograft group(P<0.05).Conclusion:Phycocyanin inhibits the secretion of inflammatory cytokines and the occurrence of EMT by regulating the STAT3/NF-κB signaling pathway,promotes tumor cell apoptosis,and inhibits the growth of transplanted tumors in nude mice.