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目的：通过肝癌凋亡细胞。DNA文库的构建，进而筛选人肝癌细胞凋亡相关基因。方法：用阿霉素诱导人肝癌HCC－9204细胞凋亡，磁珠法提取mRNA，然后将其反转录为cDNA，将合成的cDNA加上EcoR1接头，并与λgt11载体臂连接，构建为表达的cDNA文库．PCR法鉴定插入片断长度，T载体法克隆插入片断。消减杂交和点杂交法相结合筛选。DNA文库。结果：肝癌凋亡细胞CDNA文库容量为1×106pfu／μgDNA，平均片断长度为1～2kb．筛选CDNA文库得到一长度为1．5kb的DNA片段，并已测其部分序列。结论：构建的肝癌凋亡细胞CDNA文库对筛选和鉴定人肝癌细胞凋亡相关基因具有重要作用。筛选的基因可能是人肝癌细胞凋亡调控基因中的一个。
Purpose: Apoptotic cells through hepatocellular carcinoma. Construction of a DNA library and screening of apoptosis-related genes in human hepatoma cells. METHODS: Apoptosis of human hepatocellular carcinoma HCC-9204 cells was induced by doxorubicin, mRNA was extracted by magnetic bead method, and then reverse transcribed into cDNA. The synthesized cDNA was coupled with EcoR1 linker and ligated with the λgt11 vector arm to construct the expression. cDNA library. The length of the insert was determined by PCR and the insert was cloned by the T-vector method. Subtractive hybridization and dot blot hybridization combined screening. DNA library. RESULTS: The apoptotic cell density of CDNA was 1×106pfu/μg DNA, and the average fragment length was 1～2kb. The CDNA library was screened to obtain a DNA fragment of 1.5 kb in length and part of its sequence was measured. CONCLUSION: The constructed CDNA library of apoptotic hepatoma cells plays an important role in the screening and identification of apoptosis-related genes in human hepatoma cells. The selected gene may be one of the apoptosis-regulating genes of human hepatoma cells.