硫酸右旋糖酐通过影响Hif-1α、VEGF的表达抑制人胃癌细胞腹腔种植转移的研究

发布时间：2018-01-17 01:02

本文关键词：硫酸右旋糖酐通过影响Hif-1α、VEGF的表达抑制人胃癌细胞腹腔种植转移的研究　出处：《宁夏医科大学》2015年硕士论文　论文类型：学位论文

更多相关文章： DS 胃癌 Hif-1α VEGF MVD值 腹腔种植转移

【摘要】：目的:探讨硫酸右旋糖酐(dextran sulfate,DS)对缺氧诱导因子(Hif-1α)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、微血管密度(MVD)的影响,观察腹腔内瘤结节种植情况,进而研究DS对人胃癌细胞腹腔种植的抑制作用及其机制。方法:将72只裸鼠分为实验组与对照组,实验组40只腹腔注射浓度为1×107/m L的BGC-823人胃癌细胞0.2 m L,24h后腹腔注射0.3%DS1m L;对照组32只于注射相同浓度细胞0.2 m L 24h后,腹腔注射生理盐水1m L。分别于1、3、7、14d时颈椎脱位法将其处死,剖腹观察腹腔内瘤结节的转移情况,应用免疫组化法检测Hif-1α、VEGF、CD34的表达,RT-PCR及Western-blot检测Hif-1α、VEGF m RNA及蛋白的表达。结果:1.人胃癌细胞种植后1天对照组与实验组均未观察到肿瘤结节,随着时间的推移,两组瘤结节数均增多,对照组增加的幅度明显高于实验组,对照组与实验组之间结节个数有显著差异(P0.001)。2.免疫组化结果显示:1天时两组之间Hif-1α的表达无差异(P0.05),3、7、14天时两组Hif-1α的表达均增加且有明显差异(P0.01);对照组与实验组VEGF的表达均增加,两组之间的表达有明显差异(P0.001),实验组明显低于对照组;早期随着时间的推移MVD值明显增高,但在后期,MVD值反而下降,总体来看,对照组与实验组之间在不同时间点上MVD值有显著差异(P0.01),实验组的MVD值明显低于对照组。3.RT-PCR结果显示:随着时间的推移,对照组与实验组Hif-1αm RNA、VEGF m RNA的表达均增加,两组之间的表达均有明显差异(P0.001),实验组明显低于对照组。4.western-blot结果显示:人胃癌细胞种植后1天对照组与实验组均未检测到Hif-1α及VEGF的表达,随着时间的推移表达增加,Hif-1α及VEGF的表达均在两组之间均有明显差异(P0.001),对照组明显高于实验组。5.随着时间的推移,对照组和实验组Hif-1α、VEGF的表达均增加,且两者之间的表达呈正相关。结论:1.DS可以抑制人胃癌细胞在腹腔内的种植转移。2.随着人胃癌细胞种植后时间的推移,DS可以下调Hif-1α、VEGF的表达及MVD值。3.大网膜肿瘤组织内,Hif-1α与VEGF、VEGF及MVD值呈正相关。4.DS可能是通过下调Hif-1α、VEGF的表达,并影响MVD值从而抑制人胃癌细胞的腹腔种植转移。
[Abstract]:Objective: to investigate the effect of dextran sulfate1 DS on hypoxia inducible factor (HIF-1 伪). Effects of vascular endothelial growth factor (VEGF) on vascular endothelial growth factor (VEGF) and microvessel density (MVD). To investigate the inhibitory effect of DS on intraperitoneal implantation of gastric cancer cells and its mechanism. Methods: 72 nude mice were divided into experimental group and control group. In the experimental group, 40 BGC-823 cells with concentration of 1 脳 10 7 / mL were injected intraperitoneally with 0.3 DS 1m L for 24 h. 32 rats in the control group were killed by cervical dislocation at the time of 14 days after the same concentration of 0.2 mL cells were injected intraperitoneally with normal saline (1m L). The metastasis of tumor nodules in abdominal cavity was observed by laparotomy, and the expression of Hif-1 伪 VEGF CD34 was detected by immunohistochemical method. Hif-1 伪 was detected by RT-PCR and Western-blot. Expression of VEGF m RNA and protein. Results 1 day after implantation of human gastric cancer cells, no tumor nodules were observed in both the control group and the experimental group, and the number of tumor nodules increased in both groups with the passage of time. The increase in the control group was significantly higher than that in the experimental group. There was significant difference in the number of nodules between the control group and the experimental group. The immunohistochemical results showed that there was no difference in the expression of Hif-1 伪 between the two groups on day 1. On the 14th day, the expression of Hif-1 伪 in the two groups increased and there was significant difference between the two groups (P 0.01). The expression of VEGF in the control group and the experimental group was increased, and the expression of P0.001 was significantly different between the two groups, and the expression of VEGF in the experimental group was significantly lower than that in the control group. In the early stage, the MVD value increased obviously with the passage of time, but decreased in the later stage. In general, there was a significant difference in MVD between the control group and the experimental group at different time points (P0.01). The MVD value of the experimental group was significantly lower than that of the control group. 3. The results of RT-PCR showed that the Hif-1 伪 m RNA of the control group and the experimental group was higher than that of the control group. The expression of VEGF m RNA increased, and there was significant difference between the two groups (P 0.001). The results of Western blot showed that the expression of Hif-1 伪 and VEGF was not detected in the control group and the experimental group 1 day after implantation. The expression of Hif-1 伪 and VEGF increased with the passage of time in both groups. The expression of Hif-1 伪 and VEGF in the control group was significantly higher than that in the experimental group. The expression of Hif-1 伪 -VEGF was increased in both control group and experimental group. Conclusion: 1. DS can inhibit the implantation metastasis of human gastric cancer cells in abdominal cavity. 2. With the passage of time after implantation of human gastric cancer cells, DS can down-regulate Hif-1 伪. VEGF expression and MVD value. 3. Hif-1 伪 was positively correlated with VEGF and MVD in omental tumors. 4. DS may be by down-regulating Hif-1 伪. The expression of VEGF and the effect of MVD on the metastasis of human gastric cancer cells.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.2

【共引文献】