發(fā)布時間：2018-06-30 06:46

  本文選題：淋巴瘤 + MALT1�。� 參考：《貴州醫(yī)科大學》2017年碩士論文

【摘要】：目的:通過觀察粘膜相關淋巴組織淋巴瘤易位基因1(mucosa-associated lymphoid tissue lymphoma translocation gene 1,MALT1)抑制劑Z-VRPR-FMK對彌漫大B細胞淋巴瘤(Diffuse large B cell lymphoma,DLBCL)裸鼠移植瘤生長的影響和核因子κB(Nuclear factor kappa B,NF-κB)通路相關的信號分子異常,探討MALT1在DLBCL生長中的作用,為發(fā)現(xiàn)DLBCL治療的新靶點提供理論依據(jù)。方法:培養(yǎng)生發(fā)中心細胞樣彌漫大B細胞淋巴瘤(germinal center B cell like-Diffuse large B cell lymphoma,GCB-DLBCL)和活化后B細胞樣彌漫大B細胞淋巴瘤(Activated B cell like-Diffuse large B cell lymphoma,ABC-DLBCL)細胞株OCI-LY1和OCI-LY10,建立DLBCL裸鼠移植瘤模型;將成瘤裸鼠隨機分為對照組和抑制劑組。抑制劑組用Z-VRPR-FMK、對照組用等量生理鹽水腹腔注射。定期測量并記錄裸鼠體重和腫瘤體積,繪制移植瘤生長曲線圖;實時熒光定量PCR(Real time PCR)檢測p65基因在核酸水平的表達,免疫組化染色檢測P65蛋白表達,蛋白印記和免疫組織化學染色檢測MALT1、A20、MMP2和MMP9蛋白的表達。對上述檢查結果進行統(tǒng)計學分析。結果:1.裸鼠成瘤:DLBCL細胞總成瘤率68%(17/25),其中OCI-LY1細胞株成瘤率為60%(9/15),成瘤的時間為18.16±3.36天,OCI-LY10細胞株成瘤率為80%(8/10),成瘤的時間為13.63±3.02天。2.裸鼠和移植瘤的生長:OCI-LY1細胞株荷瘤鼠對照組和抑制劑組之間體重和瘤塊體積變化都沒有統(tǒng)計學差異,(P均大于0.05)。兩組OCI-LY10細胞移植瘤鼠體重在處理后逐漸上升,對照組在第9天開始逐漸下降,而抑制劑組體重則持續(xù)上升,在第11天和第13天兩組體重有統(tǒng)計學差異,P分別為0.028和0.016。3.P65表達:OCI-LY1細胞移植瘤中,抑制劑組p65的mRNA和蛋白核表達均低于對照組,但差異無統(tǒng)計學意義(P=0.060),OCI-LY10細胞移植瘤中p65核酸和蛋白表達水平都較對照組顯著降低(P=0.000,P=0.028)。4.MALT1、A20、MMP2和MMP9蛋白表達:免疫組化染色顯示MALT1,A20,MMP2和MMP9陽性信號均定位于細胞質。兩種DLBCL移植瘤的抑制劑處理組MALT1,MMP2,MMP9蛋白陽性表達信號較對照組強,A20表達則相反。Western blot檢測和分析顯示:兩組OCI-LY1細胞株移植瘤比較:在抑制劑組MALT1蛋白表達顯著降低(P=0.013),而A20蛋白表達則升高(P=0.005),兩組間MMP9、MMP2表達水平無明顯差異(P=0.234,P=0.071);兩組OCI-LY10細胞移植瘤相比:在抑制劑組MALT1、MMP2、MMP9蛋白表達水平均顯著較低(P依次為0.042,0.002,0.006),A20蛋白表達顯著升高(P=0.000)。結論:1.ABC樣DLBCL中存在MALT1蛋白的異常表達,并與腫瘤中NF-κB的持續(xù)活化密切相關,MALT1可能成為ABC樣DLBCL治療的有效靶點。2.Z-VRPR-FMK能有效地抑制DLBCL中MALT1蛋白的表達,可能通過解除其對A20蛋白的裂解作用,從而抑制NF-κB的活化。3.ABC樣DLBC中,MMP2和MMP9是NF-κB活化后上調表達的靶分子,其表達影響腫瘤的生長。
[Abstract]:Objective: to investigate the effects of Z-VRPR-FMK, a translocation gene 1 (mucosa-associated lymphoid tissue lymphoma translocation gene 1) -MALT1 inhibitor, on the growth of diffuse large B cell lymphomatous lymphoma (DLBCL) xenografts in nude mice and the correlation between nuclear factor kappa B (NF- 魏 B) pathway and nuclear factor 魏 B (NF- 魏 B) pathway. Abnormal signal molecules, To explore the role of MALT1 in the growth of DLBCL and to provide theoretical basis for finding new targets for DLBCL therapy. Methods: OCI-LY1 and OCI-LY10 were cultured from germinal center cell-like diffuse large B-cell lymphoma (germinal center B cell like-Diffuse large B cell lymphoma1) and Activated B cell like-Diffuse large B cell lymphoma1 (OCI-LY1) and OCI-LY10. Nude mice were randomly divided into control group and inhibitor group. The inhibitor group was treated with Z-VRPR- FMK and the control group with the same amount of normal saline intraperitoneal injection. The body weight and tumor volume of nude mice were measured and recorded regularly, the growth curve of transplanted tumor was drawn, the expression of p65 gene at nucleic acid level was detected by Real time PCR, and the expression of p65 protein was detected by immunohistochemical staining. Protein imprinting and immunohistochemical staining were used to detect the expression of MMP2 and MMP9. The results were statistically analyzed. The result is 1: 1. The tumorigenic rate of OCI-LY1 cell line was 60% (9 / 15), the tumorigenesis time of OCI-LY10 cell line was 18.16 鹵3.36 days (80%, 8 / 10), and the tumorigenic time of OCI-LY10 cell line was 13.63 鹵3.02 days, and the tumorigenic rate of OCI-LY10 cell line was 68% (17 / 25). The tumorigenic rate of OCI-LY1 cell line was 60% (9 / 15), and the tumorigenic time was 18.16 鹵3.36 days. There was no significant difference in body weight and tumor volume between the control group and the inhibitor group (P > 0.05). The weight of OCI-LY10 transplanted tumor mice gradually increased after treatment, the control group began to decrease gradually on the 9th day, while the weight of the inhibitor group continued to increase. On the 11th and 13th days, there were significant differences in body weight between the two groups (P = 0.028 and 0.016.3.P65, respectively). The expression of p65 mRNA and protein in the inhibitor group was lower than that in the control group. The expression of p65 nucleic acid and protein in OCI-LY10 cells was significantly lower than that in the control group (P0. 000). 4. The expression of MMP2 and MMP9 protein in MALT1A20 MMP2 and MMP9 protein: the positive signals of MALT1A20 MMP2 and MMP9 were localized in cytoplasm by immunohistochemical staining. The positive expression of MALT1 mMP2mMP9 protein in the two DLBCL transplanted tumor groups was significantly lower than that in the control group. Western blot analysis showed that the expression of MALT1 protein was significantly lower in the inhibitor group than that in the control group (Pn0.013), and the expression of MALT1 protein in the two groups was significantly lower than that in the OCI-LY1 cell line. However, the expression of A20 protein increased (P0. 005), and there was no significant difference between the two groups in the expression of MMP9 and MMP2 (P0. 234, P0. 071). The expression of A20 protein in OCI-LY10 cells was significantly lower than that in the inhibitor group (P = 0. 042, 0. 002, 0. 006, P = 0. 0000), and the expression of A20 protein in OCI-LY10 cells was significantly lower than that in the control group (P = 0. 042, 0. 002P = 0. 0000). Conclusion: 1. There is abnormal expression of MALT1 protein in ABC-like DLBCL, and it is closely related to the continued activation of NF- 魏 B in tumor. MALT1 may be an effective target for ABC-like DLBCL treatment. Z-2.VRPR-FMK can effectively inhibit the expression of MALT1 protein in DLBCL. It is possible to inhibit the activation of NF- 魏 B by removing the cleavage of A20 protein. 3. MMP2 and MMP9 in ABC-like DLBC are the target molecules that up-regulate the expression of NF- 魏 B, and their expression affects the growth of tumor.
【學位授予單位】：貴州醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R733.1

【參考文獻】

相關期刊論文 前10條

1 呂紅瓊;謝玲;倪明立;;肺鱗癌中磷脂酶Cε和核因子-κBp65的表達及意義[J];中國老年學雜志;2016年22期

2 李楊;宋軍民;李曉晗;李巖;;人胃癌組織中NF-κB-p65的表達及臨床意義[J];胃腸病學和肝病學雜志;2016年11期

3 曹鐘元;趙振凱;相庚;倪清蓉;才延輝;席宇菁;趙晶;閻博;;人胃癌裸鼠原位移植瘤模型的建立及其活體熒光成像檢測[J];中國腫瘤生物治療雜志;2016年05期

4 阮妙華;王凱;王丹;周愛華;褚茂平;陳其;錢燕;;病毒性心肌炎小鼠心肌中MMP-2和NF-κB的表達[J];中國病理生理雜志;2016年09期

5 吳輝菁;于丁;楊業(yè)勤;謝蓉;湯靜;吳月兵;朱賢敏;;ABVD方案成功解救難治性彌漫大B細胞淋巴瘤1例報道[J];腫瘤防治研究;2016年11期

6 朱瑞東;馮驥良;趙大偉;李寧;;裸鼠原位肝癌移植模型的建立及其生物學觀察[J];北京醫(yī)學;2016年09期

7 楊云鵬;劉莉;李慧;;MMP-2、MMP-3和MMP-9在胃癌患者中的表達及意義[J];局解手術學雜志;2016年09期

8 王玲玲;楊文秀;李逸;陳琴;李品浩;;TNFAIP3 siRNA對彌漫大B細胞淋巴瘤OCI-LY1細胞NF-κB活性及增殖的影響[J];臨床與實驗病理學雜志;2016年04期

9 王碧玉;黃燕妮;羊文芳;;MMP-2、MMP-9在膀胱移行細胞癌組織中的表達及其臨床意義[J];海南醫(yī)學;2016年06期

10 李品浩;楊文秀;陳琴;裴媛媛;;IL-6和AG490對Burkitt淋巴瘤細胞生長的影響[J];中國病理生理雜志;2016年03期

，

本文編號：2085380