本文關(guān)鍵詞： 胰腺癌 高強度聚焦超聲 低功率 凋亡 表達譜芯片 信號通道　出處：《天津醫(yī)科大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：研究目的 評測低功率高強度聚焦超聲(high intensity focused ultra-sound, HIFU)治療人胰腺癌YY-1移植瘤的療效、安全性,探討低功率HIFU對腫瘤細胞凋亡的影響及其機制,以期為臨床應(yīng)用提供理論依據(jù)。 研究方法 1.BALB/cnu裸小鼠30只,隨機分為低功率組、高功率組、對照組,每組10只。建立人胰腺癌YY-1移植瘤裸鼠模型,并行HIFU輻照治療,治療期間觀察腫瘤體積變化、腫瘤生長速率,記錄不良反應(yīng)；各組分別于第7天、第14天收集腫瘤標本,Western blot方法分析Bax、Bcl-2表達情況,TUNEL法檢測各組腫瘤細胞凋亡率。 2.治療第7天分別收集低劑量組及對照組腫瘤組織,通過Agilent人類基因表達譜芯片檢測差異性基因表達。通過實時定量PCR(Quantitative Real time PCR)對篩選出的差異性基因進行驗證。 結(jié)果 1.治療后,低功率組、高功率組的腫瘤體積均小于對照組(P0.05),兩組間未見顯著性差異(P0.05)；在第3天、第7天、第14天,低功率組、高功率組腫瘤生長速率明顯低于對照組,具有統(tǒng)計學(xué)差異(P0.001);兩組之間比較無統(tǒng)計學(xué)差異(P0.05)。與低功率組比較,高功率組不良反應(yīng)增多,主要為皮膚燒傷和通道組織損傷(P0.05)； 2.治療后第7天,與對照組比較,低功率組、高功率組Bax均表達上調(diào),Bcl-2表達均下調(diào),有統(tǒng)計學(xué)差異(P0.05)；治療后第14天,與對照組比較,低功率組、高功率組Bax表達均進一步上調(diào),Bcl-2表達均進一步下調(diào),差異有統(tǒng)計學(xué)意義(P0.05)；TUNEL結(jié)果顯示,治療后第7天、第14天,低功率組和高功率組細胞凋亡率均高于對照組,差異有統(tǒng)計學(xué)意義(P0.05),兩組間細胞凋亡率無統(tǒng)計學(xué)差異(P0.05)。 3.表達譜芯片結(jié)果顯示,與對照組比較,低功率HIFU輻照后發(fā)生差異表達50倍以上的基因共4767個,其中上調(diào)3958個,下調(diào)809個。GO分析(Gene Ontology,GO)和pathway分析結(jié)果提示,HIFU通過上調(diào)Fas、Bid、IL1-R、Apaf-1、Caspase-8、 Caspase-9等關(guān)鍵基因表達,直接促進凋亡,還通過影響p53信號通道、MAPK信號通道、PI3K-Akt信號通道等多個信號通道中的關(guān)鍵基因間接促進腫瘤凋亡。 4.選取Caspase-8、Caspase-9、Fas、Bcl-xL行實時定量PCR驗證,結(jié)果顯示擴增良好,并與表達譜芯片結(jié)果一致。 結(jié)論： 1.低功率、高功率HIFU治療胰腺癌移植瘤均有效,高功率組局部不良反應(yīng)較多；低功率HIFU在療效不差于高功率HIFU的基礎(chǔ)上,安全性更好。 2. HIFU可上調(diào)促凋亡基因Bax的表達,下調(diào)抗凋亡基因Bcl-2的表達,并具有時間和劑量依賴關(guān)系。 3.低功率、高功率HIFU均可促進腫瘤細胞凋亡,兩組間腫瘤細胞凋亡率未見統(tǒng)計學(xué)差異。 4.低功率HIFU可通過內(nèi)源性途徑和外源性途徑促進細胞凋亡,同時,還通過影響多種信號通道、多個環(huán)節(jié)直接或間接促進腫瘤組織凋亡,并可能存在著雙向調(diào)控機制。
[Abstract]:Research purpose. To evaluate the efficacy and safety of low power high intensity focused ultrasound (HIFU) in the treatment of human pancreatic carcinoma YY-1 transplanted tumor, and to investigate the effect of low power HIFU on apoptosis of tumor cells and its mechanism, in order to provide theoretical basis for clinical application. Research method. 1. Thirty BALB / cnu nude mice were randomly divided into low power group, high power group and control group with 10 mice in each group. The nude mice model of human pancreatic carcinoma YY-1 transplanted tumor was established and treated with HIFU irradiation. The tumor volume and tumor growth rate were observed during the treatment. On the 7th and 14th day, the expression of BaxanBcl-2 was analyzed by Western blot and Tunel method was used to detect the apoptosis rate of tumor cells in each group. 2. The tumor tissues of low dose group and control group were collected on the 7th day of treatment, the differentially expressed genes were detected by Agilent human gene expression microarray, and the differentially screened genes were verified by real-time quantitative PCR(Quantitative Real time PCR. Results. 1. After treatment, the tumor volume of low power group and high power group were all smaller than that of control group (P 0.05), there was no significant difference between the two groups, and the tumor growth rate of low power group and high power group was significantly lower than that of control group on day 3, day 7 and day 14. There was no statistical difference between the two groups (P 0.05). Compared with the low power group, the adverse reactions in the high power group were more than those in the low power group, mainly in the skin burn and channel tissue injury (P0.05). 2. On the 7th day after treatment, compared with the control group, the expression of Bax in low power group and high power group were all down-regulated, with statistical difference (P 0.05), and on the 14th day after treatment, the expression of Bax in low power group was lower than that in control group. The expression of Bax was further up-regulated and the expression of Bcl-2 was further down-regulated in high power group. The results of Tunel showed that the apoptosis rate of low power group and high power group was higher than that of control group on the 7th and 14th day after treatment. The difference was statistically significant (P 0.05), and there was no significant difference in apoptosis rate between the two groups (P 0.05). 3. Compared with the control group, there were 4767 differentially expressed genes in low power HIFU irradiation, 3 958 of which were up-regulated. The down-regulation of 809 gene ontology and pathway analysis showed that HIFU could directly promote apoptosis by up-regulating the expression of caspase-8, Caspase-9 and so on. In addition, the key genes of PI3K-Akt signal pathway and other signal channels can indirectly promote tumor apoptosis by affecting p53 signal channel and MAPK signal channel. 4. Caspase-8 caspase-9 Fas-Fas-xL was selected for real-time quantitative PCR verification. The results showed that the amplification was good, and the results were consistent with the results of expression microarray. Conclusion:. 1. Low power and high power HIFU were effective in the treatment of pancreatic cancer transplanted tumor, and the local adverse reactions were more in the high power group, and the low power HIFU was more safe than the high power HIFU. 2. HIFU could up-regulate the expression of pro-apoptotic gene Bax and down-regulate the expression of anti-apoptotic gene Bcl-2 in a time-and dose-dependent manner. 3. Low power and high power HIFU could promote the apoptosis of tumor cells, but there was no statistical difference between the two groups. 4. Low power HIFU can promote apoptosis through endogenous pathway and exogenous pathway. At the same time, it can directly or indirectly promote apoptosis of tumor tissue by affecting many signal channels, and there may be bidirectional regulation mechanism.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R735.9;R445.1

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