Rh-Endostatin作用肺癌血管正常化时相与CA9表达的关系初探

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（0.2 mL/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），9 d，1次/d。对照组小鼠同时点腹腔注射生理盐水（NS） 0.2 mL/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。

英文摘要:

Objective To explore the relationship between normalization of tumor microvessels and CA9 for rh-Endostatin to inhibit Lewis lung cancer (LLC) and the expression level of CA9 in LLC. Methods Lewis cells of logarithmic growth phase were collected and made into 1×10⁶ mL^-1 cell suspensions were prepared. The transplanted tumor model of LLC was established on C57/BL6 mice by injected 0.2 mL cell suspensions/mice into 40 C57/BL6 mice. 40 LLC mice were randomly divided into control group and rh-ES group (20 mice per group). Control group experienced treatment of intraperitoneal injection (ip) for 0.2 mL NS/d, while rh-ES group was treated for 5 mg rh-ES/(kg·d) from the first to the ninth day. The samples of 5 mice were obtained from day 2, day 4, day 6 and day 9 after treatment in control group or rh-ES group, respectively. CA9 was tested by IHC in LLC and paracancerous tissues and estimated by RT-PCR and ELISA in the each time point of both rh-ES group and control group, respectively. Results The transplanted tumor model of LLC on C57/BL6 mice was established successfully. The expression of CA9 decreased on day 4 and day 6 in rh-ES group estimated by RT-PCR and ELISA, which indicated some great significance when compared with day 2, day 9 in rh-ES group and day 4, day 6 in control group (P<0.05), and the expression of CA9 in day 2, day 4, day 6, day 9 tested by IHC was higher in

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