乙醛脱氢酶1作为喉癌Hep-2细胞系肿瘤干细胞标志物的实验研究及初步临床分析

发布时间：2018-08-26 21:11

【摘要】：目的探讨乙醛脱氢酶1（aldehyde dehydrogenase1,ALDH1）在人喉癌Hep-2细胞系中的表达,观察ALDH1不同亚群肿瘤细胞的体外生长特性。方法应用荧光细胞化学染色及流式细胞术，观察和检测喉癌Hep-2细胞系中的ALDH1的表达；应用流式分选术分离ALDH1~(br)亚群细胞, MTT法检测ALDH1不同亚群细胞增殖能力，动态流式检测ALDH1~(br)亚群体外分化能力。结果喉癌Hep-2细胞系中ALDH1呈不同程度表达，其中有2.9±0.6%的细胞ALDH1呈高表达；MTT法示ALDH1~(br)亚群的增殖能力高于ALDH1low亚群和未分选细胞；在含血清的培养液中，经过6天培养，ALDH1~(br)亚群细胞由最初分选后的94.2±3.8%下降至分选前水平。结论喉癌Hep-2细胞系中,ALDH1~(br)亚群细胞比例较少，ALDH1~(br)亚群细胞具有强的体外分化和增殖能力。目的检测ALDH1不同表达亚群细胞的体内成瘤能力，确立ALDH1为喉癌Hep-2细胞系肿瘤干细胞标志物。方法将分选后的各亚群细胞进行培养，HE染色观察不同亚群细胞的生长情况；流式细胞仪检测ALDH1~(br)亚群细胞和ALDH1low亚群细胞的细胞周期分布情况；三组细胞以5×104数量级注入裸鼠肩项部皮下，观察各组的成瘤情况。结果分选后的各组细胞生长状态相似，具有肿瘤细胞基本生长形态；各组细胞的细胞周期分布无差异；ALDH1~(br)亚群细胞的致瘤时间短，肿瘤生长速度快于其他两组，具有统计学差异（P＜0.05）。结论ALDH1~(br)亚群细胞与ALDH1low亚群细胞和未分选细胞相比，具有强的成瘤能力；ALDH1可以作为喉癌Hep-2细胞系肿瘤干细胞标志物。目的探讨乙醛脱氢酶1在喉癌组织中的表达及临床病理意义。方法采用免疫组化Elivision法对2004年1月~2006年12月间我院收治的86例喉癌患者的喉癌组织及其正常喉组织进行ALDH1检测，结合临床病理特征进行相关性分析。结果ALDH1在喉正常组织中不表达或低表达，在喉癌组织中不同程度表达，其中高表达25例，，ALDH1的表达与患者年龄、喉癌的部位、浸润范围、临床分期无明显相关性（P0.05）；有淋巴结转移患者ALDH1表达高于淋巴结阴性患者，具有统计学意义（P0.05）；5年生存率ALDH1阳性患者明显低于ALDH1阴性患者（P0.05）。结论ALDH1在喉癌组织中的高表达与淋巴结转移有关，且可以作为喉癌预后的重要因素，对喉癌预后进行评价。
[Abstract]:Objective to investigate the expression of acetaldehyde dehydrogenase 1 (aldehyde dehydrogenase1,ALDH1) in human laryngeal carcinoma Hep-2 cell line and to observe the growth characteristics of different ALDH1 subsets in vitro. Methods fluorescent cytochemical staining and flow cytometry were used to observe and detect the expression of ALDH1 in Hep-2 cell line of laryngeal carcinoma, and ALDH1~ (br) subsets were isolated by flow sorting, and the proliferative ability of different subsets of ALDH1 was detected by MTT assay. The extracellular differentiation ability of ALDH1~ (br) subpopulations was detected by dynamic flow cytometry. Results the expression of ALDH1 was different in Hep-2 cell line of laryngeal carcinoma, among which 2.9 卤0.6% of the cells showed high expression of ALDH1. The proliferation ability of ALDH1~ (br) subsets was higher than that of ALDH1low subsets and unsorted cells, and in the culture medium containing serum, the proliferation ability of ALDH1~ (br) subsets was higher than that of unsorted cells. After 6 days of culture, ALDH1 ~ (br) subsets decreased from 94.2 卤3.8% to pre-sorting level. Conclusion the ratio of ALDH1 ~ (br) subsets in laryngeal carcinoma Hep-2 cell lines is less than that of ALDH1 ~ (br) subsets, which has strong ability of differentiation and proliferation in vitro. Objective to detect the tumorigenic ability of different expression subsets of ALDH1 and to establish ALDH1 as a marker of tumor stem cells in Hep-2 cell line of laryngeal carcinoma. Methods the cell growth of different subsets was observed by HE staining and the cell cycle distribution of ALDH1~ (br) subgroup and ALDH1low subgroup were detected by flow cytometry. Three groups of cells were injected subcutaneously into shoulder and nape of nude mice with the order of 5 脳 10 ~ 4 to observe the tumorigenesis of each group. Results the cells in each group were similar in growth state and had the basic growth morphology of tumor cells, and the cell cycle distribution of each group was not different. The tumorigenic time of ALDH1 ~ (br) subgroup cells was shorter, and the tumor growth rate was faster than that of the other two groups. There was statistical difference (P < 0.05). Conclusion compared with ALDH1low subsets and unsorted cells, ALDH1~ (br) subsets have strong tumorigenic ability. ALDH1 can be used as a marker of tumor stem cells in Hep-2 cell line of laryngeal carcinoma. Objective to investigate the expression of acetaldehyde dehydrogenase 1 in laryngeal carcinoma and its clinicopathological significance. Methods from January 2004 to December 2006, 86 cases of laryngeal carcinoma and their normal laryngeal tissues were detected by immunohistochemical Elivision method. The correlation was analyzed with clinicopathological features. Results there was no or low expression of ALDH1 in normal laryngeal tissues, but no significant correlation was found between the expression of ALDH1 in 25 cases of laryngeal carcinoma and age, the location of laryngeal carcinoma, the extent of invasion, and the clinical stage (P0.05). The expression of ALDH1 in patients with lymph node metastasis was significantly higher than that in patients with negative lymph nodes (P0.05), and the 5-year survival rate in patients with positive ALDH1 was significantly lower than that in patients with negative ALDH1 (P0.05). Conclusion the high expression of ALDH1 in laryngeal carcinoma is related to lymph node metastasis and can be used as an important prognostic factor to evaluate the prognosis of laryngeal carcinoma.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.65

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