MicroRNA-155对喉鳞癌细胞株Hep-2恶性行为的影响
发布时间：2018-10-15 13:07
【摘要】：目的：探讨：microRNA-155(miR-155)过表达对喉鳞状细胞癌细胞株Hep-2增殖、迁移及凋亡方面的影响及其可能的作用机制。 方法： 1.将miR-155模拟物(miR-155mimics)瞬时转染至Hep-2细胞,以无义序列转染组(NC组)为阴性对照,以未经任何转染处理空白转染组(Blank组)为空白对照。 2.荧光显微镜观察、测量转染效率。Real-time PCR检测不同转染组Hep-2细胞转染后miR-155表达水平。 3.CCK-8法和平板克隆形成实验评价不同转染组Hep-2细胞的增殖能力。 4.流式细胞术及Hoechst33342-PI双染法检测不同转染组LHep-2细胞的凋亡情况。 5.划痕愈合试验检测不同转染组LHep-2细胞的迁移能力。 6.统计学分析应用SPSS13.0统计软件,采用x±s描述定量资料,两组间比较采用t检验,多组间比较采用单因素方差分析(one-way ANOVA), P0.05认为差异有统计学意义。 结果： 1.荧光显微镜下观察计数显示,转染24h后,miR-155mimics和无义序列转染入Hep-2细胞,其转染率均可达90%。 2. Real-time PCR检测不同转染组转染24h后miR-155相对表达水平,miR-155组分别与Blank(?)及NC组相比miR-155表达明显增加(P0.05),约16倍。 3.CCK-8法显示1niR-155组细胞增殖能力更强,并且随着转染后培养时间的延长,细胞增殖速度更加明显(P0.05)。 4.平板克隆形成实验结果miR-155(?)且细胞集落数(832+49)明显高于Blank组(496+35)和NC组(488+39),差异具有统计学意义(P0.05)。 5.流式细胞术结果miR-155组细胞总凋亡率(0.139±0.022)明显低于Blank组(0.139±0.022)和NC组(49.23±11.21),差异具有统计学意义(P0.05)。 6. Hoechst33342-PI双染法结果示miR-155组细胞凋亡和坏死比例明显降低。 7.划痕愈合试验显示划痕48h后,miR-155组细胞愈合率(83.14±12.73)明显高于Blank组(43.31+9.69)和NC组(488+39),差异具有统计学意义(P0.05)。 结论：过表达miR-155可以促进喉鳞癌Hep-2细胞的增殖与迁移愈合能力,这可能与其抗凋亡作用有关。miR-155可能在喉鳞癌的发生发展中通过多种途径作为癌基因发挥功能。
[Abstract]:Aim: to investigate the effect of microRNA-155 (miR-155) overexpression on the proliferation, migration and apoptosis of laryngeal squamous cell carcinoma cell line Hep-2 and its possible mechanism. Methods: 1. MiR-155 mimics (miR-155mimics) were transiently transfected into Hep-2 cells. The Hep-2 cells were transfected with nonsense sequence transfection group (NC group) as negative control, and blank transfection group without any transfection treatment (Blank group) as blank control. 2. Fluorescence microscope was used to observe the transfection efficiency. Real-time PCR was used to detect the level of miR-155 expression after transfection of Hep-2 cells in different transfection groups. The proliferation ability of Hep-2 cells in different transfection groups was evaluated by 3.CCK-8 assay and plate clone formation assay. 4. Flow cytometry and Hoechst33342-PI double staining were used to detect the apoptosis of LHep-2 cells in different transfection groups. The migration ability of LHep-2 cells in different transfection groups was detected by scratch healing test. 6. 6. Statistical analysis using SPSS13.0 statistical software, using x 卤s to describe the quantitative data, two groups of comparison using t test, multi-group comparison using single factor analysis of variance (one-way ANOVA), P0.05 that the difference is statistically significant. Results: 1. After 24 hours of transfection, miR-155mimics and nonsense sequence were transfected into Hep-2 cells, and the transfection efficiency was 90. 2%. Real-time PCR was used to detect the relative expression of miR-155 in different transfection groups 24 hours after transfection, miR-155 group and Blank (?) Compared with NC group, the expression of miR-155 was significantly increased (P0.05), about 16 times. 3.CCK-8 assay showed that the proliferation ability of 1niR-155 group was stronger, and with the extension of culture time after transfection, the cell proliferation rate was more obvious (P0.05). MiR-155 (?) The number of cell colonies (832.49) was significantly higher than that of Blank (496.35) and NC (488.39), and the difference was statistically significant (P0.05). Flow cytometry showed that the total apoptosis rate in miR-155 group (0.139 卤0.022) was significantly lower than that in Blank group (0.139 卤0.022) and NC group (49.23 卤11.21), the difference was statistically significant (P0.05). The results of Hoechst33342-PI double staining showed that the percentage of apoptosis and necrosis was significantly decreased in miR-155 group. Scratch healing test showed that the cell healing rate of miR-155 group (83.14 卤12.73) was significantly higher than that of Blank group (43.31.9.69) and NC group (48839) after 48 h scratch (P0.05). Conclusion: overexpression of miR-155 can promote the proliferation and migration of Hep-2 cells, which may be related to its anti-apoptosis effect. MiR-155 may play a role as a oncogene in the pathogenesis and development of laryngeal squamous cell carcinoma.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R739.65
本文编号：2272645
[Abstract]:Aim: to investigate the effect of microRNA-155 (miR-155) overexpression on the proliferation, migration and apoptosis of laryngeal squamous cell carcinoma cell line Hep-2 and its possible mechanism. Methods: 1. MiR-155 mimics (miR-155mimics) were transiently transfected into Hep-2 cells. The Hep-2 cells were transfected with nonsense sequence transfection group (NC group) as negative control, and blank transfection group without any transfection treatment (Blank group) as blank control. 2. Fluorescence microscope was used to observe the transfection efficiency. Real-time PCR was used to detect the level of miR-155 expression after transfection of Hep-2 cells in different transfection groups. The proliferation ability of Hep-2 cells in different transfection groups was evaluated by 3.CCK-8 assay and plate clone formation assay. 4. Flow cytometry and Hoechst33342-PI double staining were used to detect the apoptosis of LHep-2 cells in different transfection groups. The migration ability of LHep-2 cells in different transfection groups was detected by scratch healing test. 6. 6. Statistical analysis using SPSS13.0 statistical software, using x 卤s to describe the quantitative data, two groups of comparison using t test, multi-group comparison using single factor analysis of variance (one-way ANOVA), P0.05 that the difference is statistically significant. Results: 1. After 24 hours of transfection, miR-155mimics and nonsense sequence were transfected into Hep-2 cells, and the transfection efficiency was 90. 2%. Real-time PCR was used to detect the relative expression of miR-155 in different transfection groups 24 hours after transfection, miR-155 group and Blank (?) Compared with NC group, the expression of miR-155 was significantly increased (P0.05), about 16 times. 3.CCK-8 assay showed that the proliferation ability of 1niR-155 group was stronger, and with the extension of culture time after transfection, the cell proliferation rate was more obvious (P0.05). MiR-155 (?) The number of cell colonies (832.49) was significantly higher than that of Blank (496.35) and NC (488.39), and the difference was statistically significant (P0.05). Flow cytometry showed that the total apoptosis rate in miR-155 group (0.139 卤0.022) was significantly lower than that in Blank group (0.139 卤0.022) and NC group (49.23 卤11.21), the difference was statistically significant (P0.05). The results of Hoechst33342-PI double staining showed that the percentage of apoptosis and necrosis was significantly decreased in miR-155 group. Scratch healing test showed that the cell healing rate of miR-155 group (83.14 卤12.73) was significantly higher than that of Blank group (43.31.9.69) and NC group (48839) after 48 h scratch (P0.05). Conclusion: overexpression of miR-155 can promote the proliferation and migration of Hep-2 cells, which may be related to its anti-apoptosis effect. MiR-155 may play a role as a oncogene in the pathogenesis and development of laryngeal squamous cell carcinoma.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R739.65
【参考文献】
相关期刊论文 前4条
1 王苹;付涛;王绪锐;祝威;;应用微阵列芯片分析喉鳞状细胞癌miRNA与正常黏膜表达差异的初步研究[J];临床耳鼻咽喉头颈外科杂志;2010年12期
2 钟琦;房居高;黄志刚;陈晓红;周维国;陈学军;许洪波;马泓智;;喉鳞状细胞癌组织miRNA表达的初步研究[J];中华肿瘤防治杂志;2010年14期
3 黄选东;周雪瑞;李红;茅玲;刘芳芳;;乳腺癌组织miR-155表达及其与临床病理特征关系的分析[J];中华肿瘤防治杂志;2011年22期
4 李劲松;黄洪章;宋尔卫;潘朝斌;陈伟良;武东辉;林钊宇;;舌鳞癌组织microRNA表达谱的检测和分析[J];中华口腔医学研究杂志(电子版);2008年06期
相关博士学位论文 前1条
1 魏明辉;喉鳞癌及癌旁正常组织中microRNA的差异表达分析[D];中国协和医科大学;2009年
，本文编号：2272645
本文链接：https://www.wllwen.com/yixuelunwen/yank/2272645.html
最近更新
教材专著
