细丝蛋白A的表达对恶性黑色素瘤的调节作用
发布时间：2018-11-27 16:06
【摘要】：背景：近年来黑色素瘤的发病率呈逐年上升，已成为所有恶性肿瘤中发病率增长最快的肿瘤。恶性黑色素瘤早期经外科扩大切除术后，95%可得到治愈。但是早期恶性黑色素瘤的症状与体征不明显，常被忽视，患者就诊时往往已进入肿瘤的快速生长期，或已发生血液和淋巴转移，此时手术已不能达到无瘤状态，则只能进行全身治疗。目前，MM相对敏感的化疗药物有达卡巴嗪、替莫唑胺、铂类等，然而其单药客观有效率均低于20%。达卡巴嗪是临床中治疗恶性黑色素瘤的主导药物，是美国FDA批准的治疗转移性恶性黑色素瘤的标准药物，也被视为临床试验中评估新药疗效的基准。因此，寻找一种有效的抗恶性黑色素瘤药物及治疗靶点，或有效的药物联合方案势在必行。 FLNa是肌动蛋白结合蛋白，在多种细胞中广泛表达，其可参与细胞膜受体转位、信号转导及基因转录调节等多种功能，与细胞的黏附、增殖、迁移、侵袭、肿瘤发生等过程密切相关。相关研究表明：FLNa与DNA修复蛋白BRCA2有关，当FLNa缺乏时，修复蛋白BRCA2表达减弱，DNA损伤的修复能力下降，导致细胞的DNA损伤严重，因此沉默FLNa的表达可增强细胞对DNA损伤的敏感性。因此本研究选用DTIC、VP-16等与DNA损伤有关的化疗药物作用于M2/A7细胞，研究药物对细胞的作用与FLNa的表达水平的关系，为黑色素瘤的个体化治疗寻找新的药物及新的治疗靶点。 目的：检测FLNa的表达对于黑色素瘤M2/A7细胞的影响 方法：本研究首先对于黑色素瘤M2/A7细胞系的FLNa的表达情况进行验证，之后应用MTT法检测DTIC、VP-16对于黑色素瘤M2/A7细胞系（M2-FLNa阴性表达，A7-FLNa阳性表达）的抑制率。同时用流式细胞术检测DTIC、VP-16是否能诱导黑色素瘤M2/A7细胞系的凋亡。最后采用单细胞琼脂糖凝胶电泳法检测DTIC、VP-16对于黑色素瘤M2/A7细胞系的DNA损伤。 结果：（1）DTIC及VP-16对FLNa阴性表达的M2细胞的抑制明显强于阳性表达的A7细胞。 （2）DTIC及VP-16可诱导M2及A7细胞的凋亡，，但对FLNa阴性表达的M2细胞的凋亡作用较阳性表达的A7细胞明显。 （3）DTIC及VP-16可引起M2及A7细胞DNA损伤，但对FLNa阴性表达的M2细胞DNA损伤作用更强。 结论：（1）DTIC及VP-16可诱导M2及A7细胞的凋亡，且相同浓度的药物对FLNa阴性表达的M2细胞的凋亡作用更明显。 （2）DTIC及VP-16可引起M2及A7细胞DNA损伤，且相同浓度的药物对FLNa阴性表达的M2细胞DNA损伤作用更强。 （3）FLNa阴性表达的M2细胞对于DTIC及VP-16更加敏感。
[Abstract]:Background: in recent years, the incidence of melanoma has increased year by year, and has become the fastest growing of all malignant tumors. 95% of malignant melanoma can be cured after early surgical excision. However, the symptoms and signs of early malignant melanoma are not obvious, are often ignored, the patient often has entered the rapid growth stage of the tumor, or has occurred blood and lymphatic metastasis, at this time, the operation can no longer reach a tumor-free state. Then can only carry on the whole body treatment. At present, the relatively sensitive chemotherapeutic drugs of MM are dacarbazide, temozolamide, platinum, etc. However, the objective effective rate of single drug is lower than 20%. Dacamaxine is the leading drug in the treatment of malignant melanoma. It is the standard drug approved by FDA for the treatment of metastatic malignant melanoma. It is also regarded as the benchmark for evaluating the efficacy of new drugs in clinical trials. Therefore, it is imperative to find an effective anti-malignant melanoma drug and therapeutic target, or an effective combination of drugs. FLNa is an actin binding protein widely expressed in many kinds of cells. It can participate in cell membrane receptor translocation, signal transduction, gene transcription regulation and other functions, adhesion, proliferation, migration, invasion and so on. Tumorigenesis and other processes are closely related. Related studies have shown that FLNa is related to DNA repair protein BRCA2. When FLNa is deficient, the expression of repair protein BRCA2 weakens, and the repair ability of DNA damage decreases, which leads to the serious DNA damage of cells. Therefore, silencing the expression of FLNa can enhance the sensitivity of cells to DNA damage. In this study, DTIC,VP-16 and other chemotherapeutic drugs related to DNA damage were selected to act on M2/A7 cells, and to study the relationship between the effect of drugs on cells and the expression level of FLNa. To find new drugs and new therapeutic targets for individualized treatment of melanoma. Objective: to detect the effect of FLNa expression on melanoma M2/A7 cells. In this study, the expression of FLNa in melanoma M2/A7 cell line was verified firstly, and then DTIC, was detected by MTT assay. Inhibitory rate of VP-16 on melanoma M2/A7 cell line (M2-FLNa negative expression, A7-FLNa positive expression). At the same time, flow cytometry was used to detect whether DTIC,VP-16 could induce apoptosis of melanoma M2/A7 cell line. Finally, single cell agarose gel electrophoresis was used to detect the DNA damage of melanoma M2/A7 cell line induced by DTIC,VP-16. Results: (1) the inhibition of FLNa negative M2 cells by DTIC and VP-16 was significantly stronger than that of positive A7 cells. (2) DTIC and VP-16 could induce the apoptosis of M2 and A7 cells, but the apoptosis of M2 cells with negative FLNa expression was more obvious than that of A7 cells with positive expression. (3) DTIC and VP-16 could induce DNA damage in M2 and A7 cells, but DNA damage was stronger in M2 cells with negative FLNa expression. Conclusion: (1) DTIC and VP-16 can induce the apoptosis of M2 and A7 cells, and the same concentration of drugs can induce the apoptosis of M2 cells with negative expression of FLNa. (2) DTIC and VP-16 could induce DNA damage in M2 and A7 cells, and the same concentration of drugs had stronger effect on DNA damage of M2 cells with negative FLNa expression. (3) M 2 cells with negative FLNa expression were more sensitive to DTIC and VP-16.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.5
本文编号：2361390
[Abstract]:Background: in recent years, the incidence of melanoma has increased year by year, and has become the fastest growing of all malignant tumors. 95% of malignant melanoma can be cured after early surgical excision. However, the symptoms and signs of early malignant melanoma are not obvious, are often ignored, the patient often has entered the rapid growth stage of the tumor, or has occurred blood and lymphatic metastasis, at this time, the operation can no longer reach a tumor-free state. Then can only carry on the whole body treatment. At present, the relatively sensitive chemotherapeutic drugs of MM are dacarbazide, temozolamide, platinum, etc. However, the objective effective rate of single drug is lower than 20%. Dacamaxine is the leading drug in the treatment of malignant melanoma. It is the standard drug approved by FDA for the treatment of metastatic malignant melanoma. It is also regarded as the benchmark for evaluating the efficacy of new drugs in clinical trials. Therefore, it is imperative to find an effective anti-malignant melanoma drug and therapeutic target, or an effective combination of drugs. FLNa is an actin binding protein widely expressed in many kinds of cells. It can participate in cell membrane receptor translocation, signal transduction, gene transcription regulation and other functions, adhesion, proliferation, migration, invasion and so on. Tumorigenesis and other processes are closely related. Related studies have shown that FLNa is related to DNA repair protein BRCA2. When FLNa is deficient, the expression of repair protein BRCA2 weakens, and the repair ability of DNA damage decreases, which leads to the serious DNA damage of cells. Therefore, silencing the expression of FLNa can enhance the sensitivity of cells to DNA damage. In this study, DTIC,VP-16 and other chemotherapeutic drugs related to DNA damage were selected to act on M2/A7 cells, and to study the relationship between the effect of drugs on cells and the expression level of FLNa. To find new drugs and new therapeutic targets for individualized treatment of melanoma. Objective: to detect the effect of FLNa expression on melanoma M2/A7 cells. In this study, the expression of FLNa in melanoma M2/A7 cell line was verified firstly, and then DTIC, was detected by MTT assay. Inhibitory rate of VP-16 on melanoma M2/A7 cell line (M2-FLNa negative expression, A7-FLNa positive expression). At the same time, flow cytometry was used to detect whether DTIC,VP-16 could induce apoptosis of melanoma M2/A7 cell line. Finally, single cell agarose gel electrophoresis was used to detect the DNA damage of melanoma M2/A7 cell line induced by DTIC,VP-16. Results: (1) the inhibition of FLNa negative M2 cells by DTIC and VP-16 was significantly stronger than that of positive A7 cells. (2) DTIC and VP-16 could induce the apoptosis of M2 and A7 cells, but the apoptosis of M2 cells with negative FLNa expression was more obvious than that of A7 cells with positive expression. (3) DTIC and VP-16 could induce DNA damage in M2 and A7 cells, but DNA damage was stronger in M2 cells with negative FLNa expression. Conclusion: (1) DTIC and VP-16 can induce the apoptosis of M2 and A7 cells, and the same concentration of drugs can induce the apoptosis of M2 cells with negative expression of FLNa. (2) DTIC and VP-16 could induce DNA damage in M2 and A7 cells, and the same concentration of drugs had stronger effect on DNA damage of M2 cells with negative FLNa expression. (3) M 2 cells with negative FLNa expression were more sensitive to DTIC and VP-16.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.5
【参考文献】
相关期刊论文 前2条
1 史建伟;刘会艳;王贵英;于跃明;王小玲;杨会钗;王士杰;;细丝蛋白A在结直肠腺癌中的表达及临床意义[J];中国老年学杂志;2010年13期
2 吴艳萍;李京彬;赵瑞景;王小玲;单保恩;朱铁年;;细丝蛋白A在浸润性乳腺癌中的表达及意义[J];肿瘤;2009年07期
本文编号：2361390
本文链接：https://www.wllwen.com/yixuelunwen/pifb/2361390.html
最近更新
教材专著
