Growth Mechanism of Esophageal Carcinoma Using Protein Free Culture (Relation of Oncogenes and Cell Growth)

阐述了癌基因和抑制基因与细胞培养的关系，首先，通过与c-myc、c-erbB、hst-1、k-ras、b-FGF、CyclinD 1的Southern和北方杂交，分析了23个人食管鳞状细胞癌（ESC）组织来源的细胞系的基因扩增情况。这些基因没有表现出建立细胞系的潜力，除了CyclinD 1的表达表现出轻微的潜力，建立细胞系在蛋白质自由的环境。其次，我们进行等位基因分析，以调查是否肿瘤抑制基因参与食管癌。在3 p、5 q、6p、8 p、9 p、9 q、11 p、13 q、17 p、17 q、18 q和19 q上观察到30%的信息病例的频繁杂合性丢失（洛）。其中，5 q、13 q、17 p和18 q的洛缺失被认为涉及APC、RB、p53和DCC，而染色体18 q的缺失分析显示，常见的缺失区域不包括DCC位点。60例原发性食管鳞癌的筛选 ...更多信息 对29株人ESC细胞系进行p53基因突变的检测，结果显示，尽管5号染色体上存在高频率的洛缺失，但未发现APC基因突变。在22个细胞系中发现突变（76%）。我们还检查了65个新鲜的肿瘤样本，我们试图从所有这些样本中建立细胞系，并在26个样本中检测到突变（40%）。新鲜肿瘤中p53基因的状态与细胞系的建立之间没有显著相关性。我们的结论是，p53突变是不重要的，在建立培养的人ESC细胞系，但在体外提供了一个生长优势。p53突变与无蛋白环境中细胞生长无相关性，我们还研究了ESC中p53突变、小鼠双微体2（MDM 2）扩增和人乳头瘤病毒（HPV）感染之间的关系，在大多数肿瘤中，MDM 2基因扩增或HPV感染与p53突变无关。在细胞培养中，MDM 2基因扩增仅与细胞培养有显著相关性。CyclinD 1基因扩增与患者的不良预后显著相关，我们还发现一组p53突变和/或p53突变的患者，或MDM 2扩增的细胞存活期明显缩短，我们正在研究细胞侵袭性和细胞膜流动性与蛋白质游离的关系，文化少

The relation of oncogenes and suppressor genes to cell culture was described.First, gene amplification was analyzed in 23 cell lines derived from human esophageal squamous cell carcinoma (ESC) tissues by Southern and Northern hybridizations to c-myc, c-erbB,hst-1, k-ras, b-FGF,CyclinD1. These genes showed no potential for establishing cell lines, with the exception of CyclinD1 expression which demonstated a slight potential to establish cell lines in a protein free environment.Secondly, we performed allelotype analysis to investigate whether tumor suppressor genes are involved in esophageal cancer. Frequent loss of heterozygosity (LOH) of 30%>of the informative cases was observed on chromosomes 3p, 5q, 6p, 8p, 9p, 9q, 11p, 13q, 17p, 17q, 18q, and 19q. Among these, LOH on 5q, 13q, 17p, and 18q was considered to involve the APC,RB,p53, and DCC.However, deletion analysis of chromosome 18q revealed that the region commonly lost did not include the DCC locus. Screening of 60 primary ESC tum … More ors and 20 cultured ESC cell lines for the mutation of the APC gene revealed that there was not any mutation despite the high frequency of LOH on chromosome 5q.Thirdly, we screened 29 human ESC cell lines for mutations of the p53 gene through all coding exons and non-intron junctions. Mutations were found in 22 cell lines (76%) . We also examined 65 fresh tumor samples, from all of which we tried to establish cell lines, and detected mutations in 26 samples (40%) . There was no significant correlation between the status of the p53 gene in fresh tumors and the establishment of cell lines. We conclude that p53 mutation is not important in establishing cultured human ESC cell lines but provides a growth advantage in vitro. Neither was a relationship observed between p53 mutation and cell growth in protein free environment.We also examined relationships between p53 mutation, murine double minute 2 (MDM2) amplification, and human papillomavirus (HPV) infection in ESC.In most tumors, amplification of the MDM2 gene or infection of HPV was not associated with p53 mutations. In cell culture, MDM2 gene amplification only had a significant relation to cell culture.With regard to patients prognosis, CyclinD1 gene amplification was significantly correlated to poor prognosis in patients and we also found that a group of patients with p53 mutation and/or MDM2 amplification exhibited a significantly shorter survival period.We are now studying the relationship of invasive character and cell membrane fluidity to protein free culture. Less