【作者】 陈启龙；

【导师】 聂刘旺；

【作者基本信息】 安徽师范大学 ， 动物学， 2004， 硕士

【摘要】 SOX基因是一类与DNA序列特异结合编码转录因子的基因家族,其编码的产物具有一个HMG保守基序，在个体发育过程中参与多种发育过程的调控。人类SOX基因的缺失或突变可导致发育异常和严重的先天性疾病。如人类SOX9突变可引起CD综合征和性反转，SOX10突变引起Waardenbury—Hisschsprang（WS）综合征(WS)等。石蜡包埋标本DNA对医学上大宗病例的回顾性研究具有重要意义。本文从石蜡包埋肺癌组织中，采用几种不同的预处理方法，标本组织经二甲苯脱蜡后，用PBS缓冲液60℃温浴处理4-8h，再用不同梯度酒精处理，最后用PBS缓冲液常温处理4-8h。比直接用酒精梯度处理所获得的DNA质量要高，且PBS缓冲液的pH值在 7.5-8.5时最佳。肺癌是当前常见的恶性肿瘤之一，其致病原因也非常复杂。研究表明SOX4基因的变化与肺癌的发生具有一定的关系。本文采用PCR-SSCP及PCR产物直接测序等技术，对15例肺癌患者的组织标本及8 例正常对照个体血细胞SOX4基因编码区进行了突变分析。结果在15例肺癌组织中筛选出5例SOX4基因突变，分别位于461bp、679bp、687bp、692bp、698bp、712bp、713bp和716bp处，而8 例正常对照个体中均未检出。在5例突变中，4例在692、698、713、716bp由G/C/T/C突变为C/G/C/T；2例在679、712bp由G/C突变为A/ T。1例在461bp由C突变为A；1例在687bp由G突变为C。所有突变位点除461bp外，其余均位于HMG-box内，而氨基酸突变均位于HMG盒内。提示SOX4基因突变可能与肺癌的发生有一定的关系。本研究首次报道了肺癌组织中SOX4基因突变的情况，为探索SOX基因突变与人类疾病发生之间的关系提供了分子资料。更多还原

【Abstract】 The SOX gene family is characterized by the presence of a HMG domain involved in various developmental processes. The encoded proteins are capable of binding to DNA in a sequence-specific manner. In human, the deletion or mutation of SOX proteins results in developmental defects and congenital disease. It has been shown that mutation in the human SOX9 gene cause campomelic dysphasia (CD) and autonomic sex reversal, the mutation of SOX10 gene cause Warrensburg syndrome (WS), and so on. The specimen DNA is important for the research of reviewing large quantity cases in medicine. This paper reports different pretreatment methods of paraffin embedded lung cancer tissues before DNA extraction. After taking off paraffin wax with Xylene , first infuse them into PBS buffer for 4-8 hours in water bath, the temperature was 60℃, then handle them with different concentration of alcohol. The result showed that this method can acquire long DNA fragments, and the PBS buffer at pH 7.5-8.5 gave higher yields of DNA. Lung cancer is a familiar malignancy currently, the nosogenesis is very complex. The studies indicated that the SOX4 gene may be involved in the human lung carcinogenesis. Using PCR-SSCP and PCR product direct sequencing techniques, we analysed the ORF region of SOX4 gene encoding in 15 lung cancer tissues and 8 normal controls. We found point mutation at position 461、679、687、698、712、713bp in the SOX4 gene in 5 cases, but it was not found in normal controls. All of 5 mutated cases ,including 4 cases of G/C/T/C to C/G/C/T at 692、698、713、716bp, 2 cases of G/G to A/T at 679、712b,1 case of C to A at 461bp and 1 case of G to C at 687bp. We also found the mutations were located in HMG-box except the position of 461bp, and the amino acid mutations were all located in HMG-box. It suggests that the SOX4 gene mutation may be associated with the carcinogenesis of the lung cancer. Our studies first reports the SOX4 gene mutation of the lung cancer tissue, and also provides molecular data for the study of the relations between the mutation of SOX gene and disease occurrence.更多还原