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目的：将外源性野生型ｐ５３基因导入人骨肉瘤细胞株，研究野生型ｐ５３蛋白在诱导瘤细胞凋亡中的作用。方法：用ＤＮＡ转染技术将７重组野生型ｐ５３基因ＰＭ４７质粒导入人骨肉瘤细胞株ＨＯＳ８６０３中，用ＰＣＲ技术监测外源性ｐ５３基因在瘤细胞中存在的稳定性；用免疫组化方法检测在地塞米松的诱导下瘤细胞中ｐ５３的表达情况；用相差显微镜去观察凋亡瘤细胞的形态特点；用细胞凋亡原位检测技术标记并统计凋亡的瘤细胞数。结果：成功地将重组野性型ｐ５３基因ＰＭ４７质粒导入人骨肉瘤细胞株ＨＯＳ８６０３中，在ｇｐｔ选择培养基中培养两周后，分离生长出阳性细胞克隆；经ＰＣＲ证实外源性ｐ５３基因在瘤细胞内稳定存在并可随瘤细胞分裂而传给子代；当向培养液中加入１μｍｏｌ／Ｌ地塞松（Ｄｅｘ）后，８ｈ即可用免疫组化方法检测到ｐ５３蛋白的表达并可观察到瘤细胞出现凋亡的一系列形态学改变，转染后的瘤细胞凋亡率高达９０％。结论：当人骨肉瘤细胞株中重组野生型ｐ５３基因诱导过表达时，瘤细胞呈现凋亡，本实验为骨肉瘤的基因治疗提供了理论依据。
OBJECTIVE: To introduce exogenous wild-type p53 gene into human osteosarcoma cell lines to study the role of wild-type p53 protein in inducing apoptosis of tumor cells. Methods: The recombinant wild type p53 gene PM47 plasmid was introduced into human osteosarcoma cell line HOS8603 by DNA transfection technique. The stability of exogenous p53 gene in tumor cells was monitored by PCR and detected by immunohistochemistry. The expression of p53 in tumor cells was induced by dexamethasone. The morphological characteristics of apoptotic tumor cells were observed by phase contrast microscopy. The number of apoptotic tumor cells was marked and counted by in situ detection of apoptosis. Results: The recombinant wild type p53 gene PM47 plasmid was successfully introduced into human osteosarcoma cell line HOS-8603. Two weeks after culture in gpt selection medium, positive cell clones were isolated and isolated; PCR confirmed that the exogenous p53 gene was in the tumor. The cells exist stably and can be transmitted to progeny with the division of tumor cells; when 1 μmol/L of Dexone (Dex) is added to the culture fluid, the expression of p53 protein can be detected by immunohistochemistry after 8 hours and can be observed. A series of morphological changes of apoptosis appeared in tumor cells, and the apoptosis rate of tumor cells after transfection was as high as 90%. Conclusion: When the recombinant wild-type p53 gene is over-expressed in human osteosarcoma cell lines, the tumor cells exhibit apoptosis. This experiment provides a theoretical basis for gene therapy of osteosarcoma.