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PAPSS2与人子宫内膜癌细胞顺铂耐药的研究
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Abstract:
目的：探讨磷酰硫酸合成酶2 (3’-phosphoadenosine-5’-phosphosulfate synthase 2, PAPSS2)基因与人子宫内膜癌(Endometrial Cancer, EC)细胞顺铂耐药的关系。方法：qRT-PCR法检测人I型子宫内膜癌细胞ISHIKAWA及II型子宫内膜癌细胞HEC-1-B细胞中PAPSS2的表达；采用siRNA转染方法使I、II型子宫内膜癌细胞低表达PAPSS2，qRT-PCR观察转染细胞内PAPSS2表达情况；平板克隆及CCK-8法分别检测低表达PAPSS2组、空载体组及空白对照组I型及II型子宫内膜癌细胞增殖活性及顺铂半数抑制浓度(IC50)；免疫组化检测I型及II型子宫内膜癌及正常内膜组织中PAPSS2表达水平。结果：qRT-PCR结果表明，ISHIKAWA细胞中PAPSS2 mRNA水平中的表达明显高于II型子宫内膜癌细胞HEC-1-B细胞；经过siRNA转染处理后，PAPSS2低表达组PAPSS2 mRNA水平相较于空载体组及空白对照组子宫内膜癌细胞显著下降(P < 0.01)；PAPSS2-siRNA作用48 h后，两株细胞增殖显著增强，且对顺铂敏感性降低(P < 0.01)；免疫组化结果显示PAPSS2在子宫内膜癌中呈阳性表达，且癌组织中PAPSS2表达显著低于正常子宫内膜组织。结论：PAPSS2与子宫内膜癌细胞分型及顺铂耐药相关，且可能在子宫内膜癌的发生发展中发挥抑癌基因的作用。
Objective: To investigate the effects of PAPSS2 (3’-phosphoadenosine-5’-phosphosulfate synthase 2, PAPSS2) gene on cisplatin resistance in human endometrial cancer cells. Methods: The expression of PAPSS2 in human ISHIKAWA (type I endometrial cancer cells) and HEC-1-B (type II endometrial cancer cells) were detected by qRT-PCR. PAPSS2 knockdown expression vectors were constructed in the two types of cells by siRNA. The expression of PAPSS2 in transfected cells were detected by qRT-PCR. Cell proliferation and the half inhibitory concentration of cisplatin (IC50) in the PAPSS2 knockdown vector of ISHIKAWA and HEC-1-B cell lines, the empty vectors of the two cell lines and normal cell lines were assessed by plate clone formation assay and CCK-8 assay respectively. Immunohistochemistry was used to analyze the expression of PAPSS2 in the two types of endometrial cancer tissues. Results: The results of qRT-PCR show that the expression of PAPSS2 mRNA in ISHIKAWA cells is significantly higher than that of type II endometrial cancer cells HEC-1-B cells; after siRNA transfection treatment, the level of PAPSS2 mRNA in the low expression group is significantly decreased compared with the empty carrier group and negative control (P < 0.01). After 48 hours of PAPSS2-siRNA, the cell proliferation of the two cell lines was significantly enhanced, and the sensitivity to cisplatin was reduced (P < 0.01); Immunohistochemistry results showed that endometrial cancer presented positive expression of PAPSS2 and the expression of PAPSS2 in endometrial cancer tissues was lower than normal endometrial tissue. Conclusion: PAPSS2 is associated with cisplatin resistance of human endometrial cancer cells, and may serve as a tumor suppressor gene in human endometrial cancer.
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