青藤碱对BALB/c-nuu裸小鼠高侵袭性人结肠癌SW480移植瘤增殖及环氧合酶-2表达的影响

摘要

目的 观察青藤碱对高侵袭性人结肠癌细胞株SW 480移植瘤细胞周期及结肠瘤体组织中环氧合酶-2(COX-2)表达的影响,研究青藤碱防治结肠癌的机制.方法 先用5只免疫功能正常的BALB/c-nuu裸小鼠建立高侵袭性人结肠癌瘤源,3周后取直径约2 mm的瘤块移植于20只免疫缺陷的BALB/c-nuu2裸小鼠回盲浆膜凹龛部,1周后再随机分为结肠癌组和青藤碱组.青藤碱组按10 mL/(kg·d)灌10％青藤碱治疗30 d,结肠癌组灌0.9％氯化钠注射液对照.测量BALB/c-nuu裸小鼠结肠瘤体积、瘤质量并计算肿瘤抑制率;流式细胞仪检测结肠瘤体组织SW 480细胞在G1、G2、M和S期的细胞周期比例;免疫组化法检测结肠瘤体组织COX-2的表达,RT-PCR法检测COX-2 mRNA表达.结果 与结肠癌组比较,青藤碱组结肠瘤体积、瘤质量及S期细胞比值明显降低、G1、G2期细胞比值提高,肿瘤抑制率为(39.75％)(P＜0.05);COX-2和COX-2mRNA低表达(P＜0.05).结论 青藤碱可能在G1、G2和S期诱导SW 480细胞阻滞、抑制结肠癌细胞株SW 480细胞的有丝分裂,并通过下调COX-2表达而间接影响癌细胞的增殖,具有抗高侵袭性人结肠癌的作用.%Objective:To observe the effect of sinomenine on invasion of human colon cancer cell line SW 480 tumor cell cycle and colon tumor cyclooxygenase-2 (cyclooxygenase-2,COX-2) expression and mechanism of treating colon cancer with sinomenine.Methods:5 immunocompetent BALB/c-nuu mice were used to establish highly invasive human colon tumor models,3 weeks later,tumor block which had a diameter of 2 mm was transplanted to 20 immunodeficient BALB / c-nuu nude mice ileocecal serous concave niche,1 week later,they were randomly divided into colon cancer group and sinomenine group.Sinomenine group was treated by 10 mL/(kg·d) with 10％ sinomenine for 30 days (d),colon cancer group was given saline control.The measurement of BALB / c-nuu nude mice colon tumor volume,tumor weight and tumor inhibition rate was calculated;the colon tumor tissue SW was detected by flow cytometry in 480 cells in the cell cycle of G1,G2,M and S phase ratio;immunohistochemistry was used to detect the expression of colon tumor tissue COX-2,COX-2 mRNA expression was detected by RT-PCR.Results:Compared with colon cancer group,sinomenine group colon tumor volume,tumor cell ratio and quality of S stage decreased,G1 and G2 phase cells ratio increased,tumor inhibition rate was (39.75％) (P＜ 0.05);the expression of COX-2 and COX-2 mRNA was low (P＜ 0.05),the differences were statistically significant.Conclusion:Sinomenine can inhibit colon cancer SW 480 cells induced by SW 480 cell block in G1,G2 and S phase of mitosis,and through down-regulation of COX-2 expression,it indirectly influences the proliferation of cancer cells with high invasive human colon cancer.