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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.
[1] | Morice, P., et al. (2016) Endometrial Cancer. The Lancet (British Edition), 387, 1094-1108. https://doi.org/10.1016/S0140-6736(15)00130-0 |
[2] | Xu, Y.L., et al. (2012) Effect of Estrogen Sulfation by SULT1E1 and PAPSS on the Development of Estrogen‐Dependent Cancers. Cancer Science, 103, 1000-1009. https://doi.org/10.1111/j.1349-7006.2012.02258.x |
[3] | Jung, S.H., Lee, H.C., Yu, D.M., Kim, B.C., Park, S.M. and Lee, Y.S. (2016) Heparan Sulfation Is Essential for the Prevention of Cellular Senescence. Cell Death & Differentiation, 23, 417-429. https://doi.org/10.1038/cdd.2015.107 |
[4] | Siegel, R.L., Miller, K.D. and Jemal, A. (2018) Cancer Statistics, 2018. CA: A Cancer Journal for Clinicians, 68, 7-30. https://doi.org/10.3322/caac.21442 |
[5] | Song, R.X.-D. and Santen, R.J. (2003) Apoptotic Action of Estrogen. Apoptosis, 8, 55-60. https://doi.org/10.1023/A:1021649019025 |
[6] | Li, A.J., Baldwin, R.L. and Karlan, B.Y. (2003) Estrogen and Progesterone Receptor Subtype Expression in Normal and Malignant Ovarian Epithelial Cell Cultures. American Journal of Obstetrics & Gynecology, 189, 22-27. https://doi.org/10.1067/mob.2003.328 |
[7] | Rutherford, T., Brown, W.D., Sapi, E., Aschkenazi, S., Munoz, A. and Mor, G. (2000) Absence of Estrogen Receptor-β Expression in Metastatic Ovarian Cancer. Obstetrics & Gynecology, 96, 417-421. https://doi.org/10.1097/00006250-200009000-00018 |
[8] | Bardin, A., Hoffmann, P., Boulle, N., Katsaros, D., Vignon, F., Pujol, P., et al. (2004) Involvement of Estrogen Receptor β in Ovarian Carcinogenesis. Cancer Research, 64, 5861-5869. https://doi.org/10.1158/0008-5472.CAN-04-0552 |
[9] | Pujol, P., Rey, J.M., Nirde, P., Roger, P., Gastaldi, M., Laffarque, F. and Rochefort, H. (1998) Differential Expression of Estrogen Receptor-Alpha and-Beta Messenger RNA as a Potential Marker of Ovarian Carcinogenesis. Cancer Research, 58, 5367-5373. |
[10] | Hall, J.M. and McDonnell, D.P. (1999) The Estrogen Receptor β-Isoform (ERβ) of the Human Estrogen Receptor Modulates ERα Transcriptional Activity and Is a Key Regulator of the Cellular Response to Estrogens and Antiestrogens. Endocrinology, 140, 5566-5578. https://doi.org/10.1210/en.140.12.5566 |
[11] | Skliris, G.P., Munot, K., Bell, S.M., Carder, P.J., Lane, S., Horgan, K., et al. (2003) Reduced Expression of Oestrogen Receptor β in Invasive Breast Cancer and Its Re-Expression Using DNA Methyltransferase Inhibitors in a Cell Line Model. The Journal of Pathology, 201, 213-220. https://doi.org/10.1002/path.1436 |
[12] | Iwao, K., Miyoshi, Y., Egawa, C., Ikeda, N. and Noguchi, S. (2000) Quantitative Analysis of Estrogen Receptor-β mRNA and Its Variantsin Human Breast Cancers. International Journal of Cancer, 88, 733-736. https://doi.org/10.1002/1097-0215(20001201)88:5<733::AID-IJC8>3.3.CO;2-D |
[13] | Horvath, L.G., Henshall, S.M., Lee, C.S., Head, D.R., Quinn, D.I. and Makela, S. (2001) Frequent Loss of Estrogen Receptor-Beta Expression in Prostate Cancer. Cancer Research, 61, 5331-5335. |
[14] | Ya?ar, P., et al. (2017) Molecular Mechanism of Estrogen-Estrogen Receptor Signaling. Reproductive Medicine and Biology, 16, 4-20. https://doi.org/10.1002/rmb2.12006 |
[15] | Fuda, H., Shimizu, C., Lee, Y.C., Akita, H. and Strott, C.A. (2002) Characterization and Expression of Human Bifunctional 3’-Phosphoadenosine 5’-Phosphosulphate Synthase Isoforms. Biochemical Journal, 365, 497-504. https://doi.org/10.1042/bj20020044 |
[16] | Stelzer, C., Brimmer, A., Hermanns, P., Zabel, B. and Dietz, U.H. (2007) Expression Profile of Papss2 (3’-Phosphoadenosine 5’-Phosphosulfate Synthase 2) during Cartilage Formation and Skeletal Development in the Mouse Embryo. Developmental Dynamics, 236, 1313-1318. https://doi.org/10.1002/dvdy.21137 |
[17] | Kurima, K., Warman, M.L., Krishnan, S., et al. (1998) A Member of a Family of Sulfate-Activating Enzymes Causes Murine Brachymorphism. PNAS, 95, 8681-8685. https://doi.org/10.1073/pnas.95.15.8681 |