兰州大学机构库 >口腔医学院
RGD序列肽在口腔癌中的作用机理研究
Alternative TitleThe effect and mechanism of RGD peptide on oral squamous cell carcinoma
张洁
Subtype硕士
Thesis Advisor王丽京
2007-01-08
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Degree Discipline口腔临床医学
KeywordRGD 口腔癌 Tb 细胞 增殖 凋亡 金黄地鼠 血管形成
Abstract目的:1、利用 RGDV 体外干预人舌鳞状细胞癌脑转移亚系 Tb 细胞系, 探讨 RGDV对 Tb 细胞系增殖、凋亡的调控作用。2、利用 RGDV 对金黄地鼠颊囊癌模型进行靶向治疗,探讨 RGDV 在口腔
自发成瘤体系中的作用。方法:1、 RGDV 对舌癌细胞增殖凋亡的研究⑴ 通过 RGDV 作用于 Tb 细胞,采用 MTT 比色法、平板克隆形成实验以及流式细胞仪细胞内抗原检测法检测 PCNA 蛋白的表达,观察 RGDV 对Tb 细胞增殖的影响。⑵ 通过 RGDV 作用于 Tb 细胞系,采用 FITC-AnnexinV/PI 检测细胞凋亡、流式细胞仪检测细胞周期、 Hoechst/PI 双染法观察细胞形态、电镜观察细胞形态、 DNA 梯状电泳以及流式细胞仪细胞内抗原检测法检测 Caspase-3
蛋白的表达,观察 RGDV 对 Tb 细胞凋亡的影响。2、 RGDV 对金黄地鼠颊囊癌靶向治疗的研究建立金黄地鼠颊囊癌模型,利用RGDV 对金黄地鼠颊囊癌模型进行靶向治疗,定期观察地鼠颊囊癌肿瘤体积变化,治疗结束后取颊囊癌组织。常规包埋、切片,免疫组织化学法检测治疗后组织中 v-WF、 Brdu 的表达变化。统计分析实验结果。结果:1、 RGDV 对舌癌细胞增殖凋亡的研究⑴ RGDV 对 Tb 细胞增殖的影响: RGDV 干预组 Tb 细胞增殖速度低于Con 组,克隆形成率明显低于 Con 组(P <0.05); PCNA 表达水平明显下调。⑵ RGDV 对 Tb 细胞凋亡的影响: RGDV 组的凋亡细胞发生率明显增加,达到 88.3%与 Con 组相比有显著性差异(P<0.05); Tb 细胞复制被阻滞在S 期; DNA 电泳显示清晰的 DNA Ladder; Hoechst/PI 双染法观察到明显的凋亡细胞; Caspase-3 蛋白的含量同 Con 组相比明显增多。2、 RGDV 对金黄地鼠颊囊癌靶向治疗的研究⑴ 利用 RGDV 对金黄地鼠颊囊癌模型进行靶向治疗后,肿瘤体积的变
化结果: RGDV 组地鼠肿瘤体积较 Con 组显著缩小(P<0.05)。
⑵ 利用 RGDV 对金黄地鼠颊囊癌模型进行靶向治疗后, v-WF 的表达结果: RGDV 组和 Con 组 v-WF 均有表达,分别检测 MVD(微血管密度),RGDV 组 MVD 值显著低于 Con 组(P<0.05)。⑶ 利用 RGDV 对金黄地鼠颊囊癌模型进行靶向治疗后, Brdu 的表达结果: RGDV 组和 Con 组 Brdu 均有表达,分别检测 PI(增殖指数), RGDV组 PI 值显著低于 Con 组(P<0.05)。结论:1、应用 RGDV 干预 Tb 细胞后, Tb 细胞的增殖能力降低,提示 RGDV 抑制了 Tb 细胞的增殖能力;2、应用 RGDV 干预 Tb 细胞后, 可诱导 Tb 细胞凋亡提示细胞凋亡途径可能是 RGDV 影响肿瘤形成的重要途径, 并且 RGDV 可能通过 Caspase-3信号通路诱导 Tb 细胞凋亡;3、应用 RGDV 作用金黄地鼠颊囊癌后,抑制了肿瘤细胞的增殖,抑制了肿瘤的生长。4、应用 RGDV 作用金黄地鼠颊囊癌后,降低了金黄地鼠颊囊癌的微血管密度,说明 RGDV 对肿瘤新生血管起抑制作用。
Other AbstractPurpose: 1、Study on proliferation and apoptosis of Tb cells, to research the mechanism of RGDV on antitumor. 2、Study on the growth and angiogenesis of hamster buccal-pouch carcinoma by synthetic peptide RGDV, to research the mechanism of RGDV on antitumor. Methods: 1、Study on proliferation and apoptosis of Tb cells ⑴ The effect of RGDV on proliferation were investigated with MTT assay and colonogenic assay, and the expression of PCNA was measured with flow cytometry. ⑵ The effect of RGDV on apoptosis were measured with flow cytometry, FITC-Annexin V/PI Hoechst/PI ,electron microscopeand DNA fragmentation assay. The expression of caspase-3 protein was observed by flow cytometry. 2、Study on the growth and angiogenesis of hamster buccal-pouch carcinoma by synthetic peptide RGDV: Establishing golden HBP Ca, we used the specimens of hamster's oral buccal pouch. The specimens were embedded by paraffin traditionally.The expression of v-WF and Brdu were tested by using immunihistochhemical method. Statistically analyzed the result.Results : 1、Study on proliferation and apoptosis of Tb cells
⑴ The effect of RGDV on proliferation: The results showed that the growth and proliferation rate of Tb cell decreased in vitro after the addition of RGDV, and the expression of PCNA decreased too. ⑵ The effect of RGDV on apoptosis:The results showed the apoptotic rates increased and the expressions of caspase-3 protein increased relatively in Tb cells which were treated with RGDV. The cell number in the S phase increased, it indicated the cancer cells were blocked in the S phase. The DNA Ladder and the photo of the Hoechst/PI assay also proved the effect of RGDV on the apoptosis. We use the immunofluorescent staining of caspase-3 for flow cytometric analysis, and the expression of the group treated with RGDV increased. 2、Study on the growth and angiogenesis of hamster buccal-pouch carcinoma by synthetic peptide RGDV ⑴Blocked RGDV the effects on the model of oral carcinoma:The RGDV group tumor volume is smaller than that of Con group, after measure analyze there was significant difference between RGDV group and Con group(P<0.05).
⑵The expression results of v-WF after blocked by RGDV: v-WF were expressed in two groups, significant difference was found in RGDV group with MVD(P<0.05). ⑶The expression results of Brdu after blocked by RGDV: Brdu were expressed in two groups, significant difference was found in RGDV group
with PI(P<0.05). Conclusions: 1、RGDV could regulate the proliferation of tumor cells to inhibit the cancer growth.
2、RGDV plays a crucial role in the development of tumor by apoptosis and caspase-3 protein may be one of the important transductions in apoptosis of Tb cells. 3、RGDV prevent the growth and proliferation of golden HBP Ca. 4、RGDV play a role in tumor angiogenesis.
Pages54
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/341216
Collection口腔医学院
Affiliation
口腔医学院
First Author AffilicationSchool of Stomatology
Recommended Citation
GB/T 7714
张洁. RGD序列肽在口腔癌中的作用机理研究[D]. 兰州. 兰州大学,2007.
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