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目的初步探索重组人血管内皮抑素(rh-Endostatin,rh-ES)作用于肺癌的血管正常化时相与CA9的关系,以及CA9在肺癌中的表达水平。方法收集对数生长期的Lewis细胞,制成1×106 mL-1的单细胞悬液,注射入40只C57/BL6小鼠(0.2mL/只),建立Lewis肺癌皮下移植瘤(LLC)模型,然后随机分成对照组和rh-ES组,各20只。rh-ES组小鼠腹腔注射rh-ES 5 mg/(kg·d),9d,1次/d。对照组小鼠同时点腹腔注射生理盐水(NS)0.2mL/次。于治疗第2、4、6、9天,每组各处死5只小鼠。免疫组化检测肿瘤组织和癌旁组织中CA9表达,Real-time PCR及ELISA技术分别检测两组不同时间点肿瘤组织中CA9表达。结果 C57/BL6小鼠成瘤率为100%。同时,对照组不同时间点CA9在肿瘤组织中表达均高于癌旁组织(P<0.05)。Real-time PCR及ELISA发现在LLC移植瘤模型中CA9基因和蛋白表达在rh-ES组给药后第4天和第6天(血管正常化时相)降低,与同组第2天、第9天比较,差异有统计学意义(P<0.05),与对照组第4天和第6天比较,差异有统计学意义(P<0.05)。结论 CA9在肿瘤组织中高表达。rh-ES可于血管正常化时间段内降低CA9表达,逆转Lewis肺癌乏氧。
Objective To investigate the relationship between rh-endostatin (rh-ES) and CA9 in lung cancer and the expression of CA9 in lung cancer. Methods Lewis lung cells in logarithmic growth phase were harvested and single cell suspension (1 × 106 mL-1) was injected into 40 C57 / BL6 mice (0.2 mL / mouse) to establish a Lewis lung carcinoma subcutaneous transplantation tumor model , And then randomly divided into control group and rh-ES group, each 20. The mice in rh-ES group were injected intraperitoneally with rh-ES 5 mg / (kg · d), once a day for 9 days. The control group mice were injected intraperitoneally with saline (NS) 0.2 mL / time. On the 2nd, 4th, 6th and 9th day of treatment, 5 mice were sacrificed in each group. The expression of CA9 in tumor tissues and adjacent tissues was detected by immunohistochemistry. The expression of CA9 in tumor tissues was detected by Real-time PCR and ELISA respectively. Results The tumor formation rate of C57 / BL6 mice was 100%. Meanwhile, the expression of CA9 in the control group at different time points was higher than that in the adjacent tissues (P <0.05). Real-time PCR and ELISA showed that CA9 gene and protein expression in the LLC xenograft model was decreased on the 4th and 6th days (normalization of blood vessels) in the rh-ES group, The difference was statistically significant (P <0.05) on the 9th day compared with the control group on the 4th and 6th day, the difference was statistically significant (P <0.05). Conclusion CA9 is highly expressed in tumor tissues. rh-ES can reduce CA9 expression during the period of vascular normalization and reverse hypoxia of Lewis lung carcinoma.