肿瘤巨细胞与口腔鳞癌临床病理因素相关性研究及其相关机制初步探讨
发布时间：2019-01-04 17:34
【摘要】：目的 探究肿瘤巨细胞(giant cancer cell,GCC)在口腔鳞癌(oral squamous cell carcinoma,OSCC)组织中的存在情况及其数目与OSCC临床病理因素的关系。并初步探究肿瘤巨细胞产生的可能机制及其对OSCC增殖、耐药和迁移能力的影响。方法 1、肿瘤巨细胞在OSCC中的存在情况及其数目与临床病理因素的相关性研究:收集2005年3月至2014年12月天津医科大学肿瘤医院76例OSCC患者的石蜡切片及临床病理资料。HE染色下观察GCC与普通肿瘤细胞的形态差异,并计数GCC。免疫组织化学染色下观察GCC与普通肿瘤细胞Ki-67表达差异。分析GCC数目与OSCC临床生物学行为及预后的关系。使用SPSS18.0对收集的数据进行分析,认为P0.05为差异具有统计学意义。2、肿瘤巨细胞产生的可能机制及其对OSCC增殖、耐药和迁移能力的影响:通过CCK-8实验确定细胞乏氧的Co Cl2浓度,用Co Cl2对OSCC细胞系进行诱导处理,显微镜下观察GCC的形成及其产生子代细胞的过程。通过real-time PCR检测诱导前后细胞干性标记物mi RNA水平表达的变化;通过western blotting检测乏氧诱导因子HIF-1α和干性标记物的蛋白水平表达变化;通过CCK-8实验检测诱导前后细胞增殖能力及耐化疗药(顺铂)能力的变化;通过划痕实验检测细胞迁移能力的变化。Western blotting检测迁移相关蛋白的表达变化。结果 1、GCC存在于口腔鳞癌组织中,且其细胞核体积大或含有多个细胞核,细胞核形态不规则,可呈哑铃型、分叶状等多种不规则形态,可见多个核仁。Ki-67在OSCC胞核中部分阳性表达,在GCC细胞核中均呈阳性表达。GCC细胞数与临床分期、组织学分级、化疗及复发相关。临床分期、淋巴结转移、组织学分级、局部复发及远处转移和OS相关;临床分期、淋巴结转移、局部复发及远处转移和DFS相关。术前诱导化疗不能改善OS和DFS。GCC数目3个/HP组OS和DFS明显较GCC数目≤3个/HP组者差,GCC细胞可能是影响OSCC预后的重要因素。临床分期、局部复发和远处转移是影响OSCC患者OS的独立预后因素;临床分期、局部复发和远处转移是影响OSCC患者DFS的独立预后因素。2、以UM1为细胞模型,通过Co Cl2多次处理诱导纯化形成GCC。乏氧后,大部分普通大小的肿瘤细胞被杀死,留下纯化的GCC或者经细胞融合形成的GCC。GCC细胞体积明显较普通肿瘤细胞大,其内可含有多个细胞核,其形态多样,通过不均分裂(出芽)产生子代细胞。Real-time PCR结果显示GCC细胞及其子代细胞的干性标记物NANOG、SOX-2在mi RNA水平表达明显升高。Western blotting结果显示GCC及其子代细胞的HIF-1α及SOX-2在蛋白水平表达明显升高。诱导形成GCC后,CCK-8结果显示肿瘤细胞的增殖能力上升,耐药能力上升;划痕实验结果显示肿瘤细胞的迁移能力上升;Western blotting检测迁移相关蛋白,E-cadherin的表达下调,N-cadherin、Vimentin的表达上调。结论 1、肿瘤巨细胞细胞数与口腔鳞癌临床分期、组织学分级、诱导化疗和局部复发相关,可能是影响口腔鳞癌预后的重要因素。肿瘤巨细胞可能在口腔鳞癌的发展及耐药过程中发挥作用。2、乏氧可诱导口腔鳞癌细胞产生肿瘤巨细胞,具有肿瘤干细胞的部分特性,肿瘤巨细胞及其子代细胞干性标记物表达上调并发生EMT,其增殖、耐药和迁移能力均增强。
[Abstract]:Objective To investigate the presence of tumor giant cell (GCC) in oral squamous cell carcinoma (OSCC) and its relationship with the clinicopathological factors of OSCC. The possible mechanism of the tumor giant cell production and its effect on the proliferation, drug resistance and migration ability of OSCC were investigated. Method 1. The presence of tumor giant cells in OSCC and its correlation with the clinical and pathological factors were studied. The paraffin sections and clinical pathological data of 76 patients with OSCC from March 2005 to December 2014 were collected. The morphological differences of GCC and normal tumor cells were observed with HE staining and the GCC was counted. The expression of Ki-67 in GCC and normal tumor cells was observed with immunohistochemical staining. The relationship between the number of GCC and the clinical biological behavior and prognosis of OSCC was analyzed. The collected data was analyzed using SPSS18. 0, which was considered to have a statistical significance on the difference of P0. 05. The possible mechanism of the tumor giant cell production and its effect on the proliferation, drug resistance and migration ability of the OSCC were determined by the CCK-8 experiment. The OSCC cell line was induced with Co Cl2, and the formation of GCC and the process of producing subcell were observed under the microscope. The changes of the expression of the expression of mi-RNA in the cell-dry marker before and after induction were detected by the real-time PCR, and the changes of the expression of the protein in the HIF-1 and the dry markers were detected by western blotting, and the ability of the cell proliferation and the resistance to chemotherapy (cisplatin) was detected by CCK-8. The change of cell migration ability was detected by a scratch test. The expression of the related protein was detected by Western blotting. Results 1, GCC was present in the oral squamous cell carcinoma, and its nuclear volume is large or contains a plurality of nuclei, and the nuclear form is irregular, and can be made into a plurality of irregular shapes such as a dumbbell type, a lobule and the like, and a plurality of nucleoli can be seen. The positive expression of Ki-67 in the core of the OSCC was positive in the nucleus of the GCC. The number of GCC cells was related to clinical stage, histological grade, chemotherapy and recurrence. Clinical stage, lymph node metastasis, histological grade, local recurrence and distant metastasis and OS related; clinical stage, lymph node metastasis, local recurrence and distant metastasis and DFS correlation. The pre-operative induction of chemotherapy could not improve OS and DFS. The number of GCC 3/ HP group OS and DFS was significantly lower than that of the 3/ HP group, and GCC cells may be an important factor in the prognosis of OSCC. Clinical staging, local recurrence and distant metastasis are independent prognostic factors that affect the OS in OSCC patients; clinical staging, local recurrence and distant metastasis are independent prognostic factors that affect DFS in OSCC patients. After spent oxygen, most of the normal size of the tumor cells are killed, leaving purified GCC or the GCC formed by cell fusion. Real-time PCR results showed that the expression of NNOG and SOX-2 in the dry markers of GCC cells and its subcells increased significantly at the level of mi RNA. The results of Western blotting showed that the expression of HIF-1 and SOX-2 in GCC and its sub-cells increased significantly at the level of protein. After the formation of GCC, the results of CCK-8 showed that the proliferation ability of the tumor cells increased and the resistance to drug resistance increased; the results of the scratch test showed that the migration ability of the tumor cells increased; the expression of E-cadherin was down-regulated by Western blotting, and the expression of N-cadherin and Vimentin was up-regulated. Conclusion 1. The number of the tumor cells in the tumor is related to the clinical stage, the histological grade, the induction of chemotherapy and the local recurrence of the oral squamous cell carcinoma, which may be an important factor in the prognosis of oral squamous cell carcinoma. the tumor giant cell can play a role in the development and the drug resistance of the oral squamous cell carcinoma. Both resistance and migration were enhanced.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8
本文编号：2400580
[Abstract]:Objective To investigate the presence of tumor giant cell (GCC) in oral squamous cell carcinoma (OSCC) and its relationship with the clinicopathological factors of OSCC. The possible mechanism of the tumor giant cell production and its effect on the proliferation, drug resistance and migration ability of OSCC were investigated. Method 1. The presence of tumor giant cells in OSCC and its correlation with the clinical and pathological factors were studied. The paraffin sections and clinical pathological data of 76 patients with OSCC from March 2005 to December 2014 were collected. The morphological differences of GCC and normal tumor cells were observed with HE staining and the GCC was counted. The expression of Ki-67 in GCC and normal tumor cells was observed with immunohistochemical staining. The relationship between the number of GCC and the clinical biological behavior and prognosis of OSCC was analyzed. The collected data was analyzed using SPSS18. 0, which was considered to have a statistical significance on the difference of P0. 05. The possible mechanism of the tumor giant cell production and its effect on the proliferation, drug resistance and migration ability of the OSCC were determined by the CCK-8 experiment. The OSCC cell line was induced with Co Cl2, and the formation of GCC and the process of producing subcell were observed under the microscope. The changes of the expression of the expression of mi-RNA in the cell-dry marker before and after induction were detected by the real-time PCR, and the changes of the expression of the protein in the HIF-1 and the dry markers were detected by western blotting, and the ability of the cell proliferation and the resistance to chemotherapy (cisplatin) was detected by CCK-8. The change of cell migration ability was detected by a scratch test. The expression of the related protein was detected by Western blotting. Results 1, GCC was present in the oral squamous cell carcinoma, and its nuclear volume is large or contains a plurality of nuclei, and the nuclear form is irregular, and can be made into a plurality of irregular shapes such as a dumbbell type, a lobule and the like, and a plurality of nucleoli can be seen. The positive expression of Ki-67 in the core of the OSCC was positive in the nucleus of the GCC. The number of GCC cells was related to clinical stage, histological grade, chemotherapy and recurrence. Clinical stage, lymph node metastasis, histological grade, local recurrence and distant metastasis and OS related; clinical stage, lymph node metastasis, local recurrence and distant metastasis and DFS correlation. The pre-operative induction of chemotherapy could not improve OS and DFS. The number of GCC 3/ HP group OS and DFS was significantly lower than that of the 3/ HP group, and GCC cells may be an important factor in the prognosis of OSCC. Clinical staging, local recurrence and distant metastasis are independent prognostic factors that affect the OS in OSCC patients; clinical staging, local recurrence and distant metastasis are independent prognostic factors that affect DFS in OSCC patients. After spent oxygen, most of the normal size of the tumor cells are killed, leaving purified GCC or the GCC formed by cell fusion. Real-time PCR results showed that the expression of NNOG and SOX-2 in the dry markers of GCC cells and its subcells increased significantly at the level of mi RNA. The results of Western blotting showed that the expression of HIF-1 and SOX-2 in GCC and its sub-cells increased significantly at the level of protein. After the formation of GCC, the results of CCK-8 showed that the proliferation ability of the tumor cells increased and the resistance to drug resistance increased; the results of the scratch test showed that the migration ability of the tumor cells increased; the expression of E-cadherin was down-regulated by Western blotting, and the expression of N-cadherin and Vimentin was up-regulated. Conclusion 1. The number of the tumor cells in the tumor is related to the clinical stage, the histological grade, the induction of chemotherapy and the local recurrence of the oral squamous cell carcinoma, which may be an important factor in the prognosis of oral squamous cell carcinoma. the tumor giant cell can play a role in the development and the drug resistance of the oral squamous cell carcinoma. Both resistance and migration were enhanced.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8
【参考文献】
相关期刊论文 前3条
1 钟来平;孙坚;郭伟;涂文勇;竺涵光;季彤;胡永杰;李军;叶为民;徐立群;何悦;杨雯君;王延安;任国欣;殷秋明;蔡以理;杨溪;张陈平;张志愿;;256例局部晚期口腔癌的生存分析[J];中国肿瘤临床;2015年04期
2 张春影;卢颖;李青;毛俊;李连宏;;上皮间质转化及其相关信号通路与乳腺癌干细胞的关系[J];中华乳腺病杂志(电子版);2015年01期
3 张挚;吴江;梁冰;皇甫超申;;缺氧诱导因子-1α与乳腺浸润性导管癌上皮—间质转化的关系[J];山东医药;2012年03期
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