摘要
目的研究nm23H1基因5’端非转录区微卫星位点的微卫星不稳定(MSI)和杂合性缺失(LOH)对肺鳞癌(SLC)nm23H1蛋白表达的影响,以探讨nm23H1基因遗传不稳定性与肺鳞癌进展的关系,为肺鳞癌临床治疗和预后提供实验依据。方法采用新鲜组织标本抽提DNA,应用荧光PCR-单链构象多态性(FPCR-SSCP)方法、免疫组织化学染色法,Leica-Qwin计算机图像分析等方法,进行nm23H1基因遗传不稳定性的研究。结果在能提供信息(杂合子)的44例肺鳞癌患者中,MSI、LOH检出率和nm23H1蛋白阳性表达率分别为11.36%、25.00%和50.00%。在TNMⅠ、Ⅱ、Ⅲ期中,MSI的检出率分别为8.70%、10.00%和18.18%,而LOH分别为30.43%、20.00%和18.18%;MSI和LOH在淋巴结转移组的检出率为10.00%和20.00%,而无淋巴结转移组则为12.50%和29.17%。然而,以上各组在统计学上均无显著差异(P>0.05)。本研究中所有MSI(共5例)全部发生在分化良好(G1为2例和G2为3例)的存活患者(1年生存率)中,但MSI的检出率与肺鳞癌分化程度及患者生存状态并无统计学上的相关性(P>0.05)。nm23H1蛋白阳性表达率在TNMⅠ期(65.22%)、无淋巴结转移组(66.67%)显著高于TNMⅢ期(18.18%)、淋巴结转移组(30.00%)(P<0.05)。另外,统计学分析表明,MSI和LOH的检出率与nm23H1蛋白表达无关(P>0.05)。计算机图像定量分析显示,在各临床病理参数影响下,各组nm23H1蛋白的表达强度也没有差异(P>0.05)。结论nm23H1蛋白可能对抑制肺鳞癌淋巴结转移有重要作用。nm23H1基因5’端非转录区微卫星的MSI和LOH对nm23H1蛋白的表达无影响,也未发现其与肺鳞癌发生、发展具有相关性。
Objective To evaluate microsatellite instability （MSI） and loss of heterozygosity （LOH） of the repetitive untranslated 5＇region of the nm23H1 gene and their influence on the expression of nm23H1 in Chinese with squamous cell lung carcinoma（SLC）. Methods Techniques such as DNA extraction from tissue and fluorescence-based pelymerase chain reaction single-strand conformation pelymorphism（FPCR-SSCP） combined with an automated DNA sequencer were used to study MSI and LOH of nm23H1. Immunohistochemistry and Leica-Qwin computer imaging techniques were used to assess the expression of nm23H1 gene. Results Of these cases of informative tumors 44 of 50, the frequencies of MSI, LOH and nm23 H1 protein positive were 11.36%, 25.00% and 50.00% respectively. The frequencies of MSI were 8.70%, 10.00% and 18.18% in TNM stage Ⅰ , Ⅱ and m respectively; The frequencies of LOH were 30.43%, 20.00% and 18.18% in TNM stage, Ⅰ , Ⅱ and m respectively and the frequency of MSI and LOH were 10.00% and 20.00% in lymph node metastasis cases compared with 12.50% and 29.17% in those without metastasis. However, the frequencies of MSI and LOH showed no correlation with TNM stage and lymph node metastasis （P 〉 0.05）. All the 5 cases with MSI positive belonged to moderate （G2） or good （G1） histological grade and all were alive in a year, but the frequency of MSI was not related to tumor differentiation and survival status of patient（ P 〉 0.05）. The positive frequency of nm23H1 protein in lymph node metastasis cases （30.00%） was significantly lower than those without metastasis（66.67% ） （ P 〈 0.05）. TNM stage Ⅲ （18 18% ） also exhibited lower positive frequency of nm23H1 protein compared with stage Ⅰ （65.22%）（P 〈 0.05）. Furthermore, the frequencies of MSI and LOH were not related to the expression of nm23H1 protein（ P 〉 0.05）. There was also no difference in nm23H1 protein expression intensity analyzed by computer imaging（ P 〉 0.05）. Conclusion The increase in the amount of nm23H1 protein expression can play an important role in restraining metastasis of squamous cell lung carcinoma. The heredity instabllity（MSl and LOH） of nm23H1 gene may not be implicated in expression of the gene and pathogenesis and progression of squamons cell lung carcinoma.
出处
《解剖学报》
CAS
CSCD
北大核心
2007年第5期552-557,共6页
Acta Anatomica Sinica
基金
浙江省卫生厅课题资助(2004B199)
