Effects of Progesterone on Expressions of p57kip2 and Cyclin E in Human Endometrioid Carcinoma Cells JEC
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摘要:目的
探讨孕激素对体外培养的人子宫内膜样癌（endometrioid carcinoma, EC）JEC细胞中p57kip2、Cyclin E表达的影响。
方法分别采用高、中、低三种浓度（1×10-4、1×10-6、1×10-8 mol/L）的孕酮（progesterone, P）处理体外培养的JEC细胞，以未加药JEC细胞作为对照组，作用24、48、72 h后，分别采用光学显微镜和电子显微镜观察JEC细胞的形态变化，MTT法检测JEC细胞的生长情况，Western blot法检测JEC细胞中p57kip2、Cyclin E蛋白的表达。
结果MTT法、光学显微镜、电子显微镜结果显示，中、低浓度P可促进JEC细胞的生长（P < 0.05），高浓度P则抑制JEC细胞的生长（P < 0.05）；Western blot结果显示，P作用于JEC细胞后：（1）作用48、72 h时，随着P浓度的增加，p57kip2蛋白的表达逐渐增高（P > 0.05），Cyclin E蛋白的表达则逐渐降低（P < 0.05）；（2）同一浓度时，随着P作用时间的延长，p57kip2蛋白在高、中、低浓度组中的表达均呈递增趋势（P < 0.05），Cyclin E蛋白在中、高浓度组的表达均呈递减趋势（P < 0.05）；p57kip2和Cyclin E蛋白的表达呈负线性相关关系（P < 0.05）。
结论在EC的发生发展中，孕酮对p57kip2、Cyclin E蛋白的表达均具有一定的调控作用。
Abstract:ObjectiveTo investigate the effects of progesterone on the expressions of p57kip2 and Cyclin E in human endometrioid carcinoma(EC) cells JEC in vitro.
MethodsThe JEC cells cultured in vitro were treated with high, medium and low concentrations(1×10-4, 1×10-6 and 1×10-8mol/L) of progesterone(P). The untreated JEC cells were taken as control group. After 24, 48, 72h, the light microscope and electron microscope were used to observe the morphological changes of JEC cells. MTT was used to test the growth conditions. Western blot was used to test the protein expressions of p57kip2 and Cyclin E in JEC cells.
ResultsMTT, light microscope and electron microscope results showed medium and low concentration of progesterone could promote JEC cells growth (P < 0.05), but high concentration of progesterone could inhibit the growth of JEC cells(P < 0.05). Western blot result showed: (1) when JEC cells were treated with progesterone for 48 and 72h, the protein expression of p57kip2 was increased with the increase of progesterone concentration (P > 0.05) while the protein expression of Cyclin E was decreased (P < 0.05); (2) at the same concentration of progesterone, the protein expression of p57kip2 in the high, medium and low concentration groups were progressively increased with the extension of time (P < 0.05), while the protein expression of Cyclin E were progressively decreased in the high and medium concentration groups (P < 0.05); the positive expression rates of p57kip2 and Cyclin E were negatively linear regression relation (P < 0.05).
ConclusionIn the occurrence and development of EC, progesterone could regulate the protein expressions of p57kip2 and Cyclin E.
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Key words:
- Endometrioid carcinoma(EC) /
- JEC cells /
- Progesterone(P) /
- p57kip2 /
- Cyclin E
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图 1 MTT检测不同浓度P作用后JEC细胞的生长曲线
Figure 1 Growth curves of JEC cells after treated with different concentrations of progesterone detected by MTT
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图 2 P作用72 h时JEC细胞的形态变化(×100)
Figure 2 Morphological changes of JEC cells after treated with progesterone for 72h (×100)
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图 3 JEC细胞P作用72 h的超微结构变化
Figure 3 Ultrastructural changes of JEC cells after treated with progesterone for 72h
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图 4 不同浓度P作用JEC细胞24 h p57kip2和Cyclin E蛋白的表达
Figure 4 Protein expressions of p57kip2 and Cyclin E in JEC cells after treated with different concentrations of progesterone for 24h
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图 5 不同浓度P作用JEC细胞48 h p57kip2和Cyclin E蛋白的表达
Figure 5 Protein expressions of p57kip2 and Cyclin E in JEC cells after treated with different concentrations of progesterone for 48h
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图 6 不同浓度P作用JEC细胞72h p57kip2和Cyclin E蛋白的表达
Figure 6 Protein expressions of p57kip2 and Cyclin E in JEC cells after treated with different concentrations of progesterone for 72h
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[1] Horrée N, van Diest PJ, van der Groep P, et al. Progressive derailment of cell cycle regulators in Endometrial carcinogenesis[J]. J Clin Pathol, 2008, 61(1): 36-42. http://cn.bing.com/academic/profile?id=2074553812&encoded=0&v=paper_preview&mkt=zh-cn
[1] Horrée N, van Diest PJ, van der Groep P, et al. Progressive derail ment of cell cycle regulators in Endometrial carcinogenesis[J]. J Clin Pathol, 2008, 61(1): 36-42. [2] Ma YQ, Wang J, Liu AJ. WHO(2014) of the uterus, uterine cervix cancer, and trophoblast disease about histological classification[J]. Zhen Duan Bing Li Xue Za Zhi, 2014, 21(9): 596-7. [马亚琪, 王 昀, 刘爱军. WHO(2014)子宫体、宫颈肿瘤和滋养细胞疾病组 织学分类[J]. 诊断病理学杂志, 2014, 21(9): 596-7.] [2] 马亚琪, 王昀, 刘爱军. WHO(2014)子宫体、宫颈肿瘤和滋养细胞疾病组织学分类[J].诊断病理学杂志, 2014, 21(9): 596-7. http://www.cnki.com.cn/Article/CJFDTotal-ZDBL201409024.htm Ma YQ, Wang J, Liu AJ. WHO(2014) of the uterus, uterine cervix cancer, and trophoblast disease about histological classification[J]. Zhen Duan Bing Li Xue Za Zhi, 2014, 21(9): 596-7. http://www.cnki.com.cn/Article/CJFDTotal-ZDBL201409024.htm
[3] Lee SH, Lee JK, Jin SM, et al. Expression of cell-cycle regulators (cyclin D1, cyclin E, p27kip1, p57kip2) in papillary thyroid carcinoma[J]. Otolaryngol Head Neck Surg, 2010, 142(3): 332-7. doi: 10.1016/j.otohns.2009.10.050
[3] Lee SH, Lee JK, Jin SM, et al. Expression of cell-cycle regulators (cyclin D1, cyclin E, p27kip1, p57kip2) in papillary thyroid carcinoma[J]. Otolaryngol Head Neck Surg, 2010, 142(3): 332-7. [4] 罗祖强, 周正平, 格桑志玛, 等.子宫内膜癌及不同子宫内膜病变组织中p57kip2、ER、PR的表达及意义[J].临床与实验病理学杂志, 2013, 29(9): 982-5. http://www.cnki.com.cn/Article/CJFDTotal-LSBL201309014.htm Luo ZQ, Zhou ZP, Ge-sang ZM, et al. Expressions and significances of p57kip2, estrogen receptor, progesterone receptor in endometrial carcinoma and endometrial different lesions[J]. Lin Chuang Yu Shi Yan Bing Li Xue Za Zhi, 2013, 29(9): 982-5. http://www.cnki.com.cn/Article/CJFDTotal-LSBL201309014.htm
[4] Luo ZQ, Zhou ZP, Ge-sang ZM, et al. Expressions and significances of p57kip2, estrogen receptor, progesterone receptor in endometrial carcinoma and endometrial different lesions[J]. Lin Chuang Yu Shi Yan Bing Li Xue Za Zhi, 2013, 29(9): 982-5. [罗 祖强, 周正平, 格桑志玛, 等. 子宫内膜癌及不同子宫内膜病变 组织中p57kip2、ER、PR的表达及意义[J]. 临床与实验病理学 杂志, 2013, 29(9): 982-5.] [5] 戴忠英.子宫内膜癌的激素治疗[J].实用妇产科杂志, 2008, 24(5): 268-71. http://www.cnki.com.cn/Article/CJFDTotal-SFCZ200805006.htm Dai ZY. Hormone therapy of endometrial carcinoma[J]. Shi Yong Fu Chan Ke Za Zhi, 2008, 24(5): 268-71. http://www.cnki.com.cn/Article/CJFDTotal-SFCZ200805006.htm
[5] Dai ZY. Hormone therapy of endometrial carcinoma[J]. Shi Yong Fu Chan Ke Za Zhi, 2008, 24(5): 268-71. [戴忠英. 子宫内膜癌 的激素治疗[J]. 实用妇产科杂志, 2008, 24(5): 268-71.] [6] Han FX, Lin H, Ru L. MTT assay for detecting 5-fluorouracil chemosensitivity of human breast carcinoma cell line[J]. Nan Fang Yi Ke Da Xue Xue Bao, 2009, 29(1): 97-9. [ 韩方璇, 林慧, 汝玲. MTT法检测乳腺癌细胞株对化疗药物5-氟脲嘧啶的敏感 性[J] 南方医科大学学报, 2009, 29(1): 97-9.] [6] 韩方璇, 林慧, 汝玲. MTT法检测乳腺癌细胞株对化疗药物5-氟脲嘧啶的敏感性[J].南方医科大学学报, 2009, 29(1): 97-9. Han FX, Lin H, Ru L. MTT assay for detecting 5-fluorouracil chemosensitivity of human breast carcinoma cell line[J]. Nan Fang Yi Ke Da Xue Xue Bao, 2009, 29(1): 97-9.
[7] 陈红平, 张垒, 康路妹, 等.雌激素对大鼠腺垂体中瘦素长型受体表达的影响[J].解剖学杂志, 2008, 31(6): 755-7. http://www.cnki.com.cn/Article/CJFDTOTAL-JPXZ200806012.htm Chen HP, Zhang L, Kang LM, et al. Effect of estrogen on the expression of long leptin receptor in rat adenohypophysis[J]. Jie Pou Xue Za Zhi, 2008, 31(6): 755-7. http://www.cnki.com.cn/Article/CJFDTOTAL-JPXZ200806012.htm
[7] Chen HP, Zhang L, Kang LM, et al. Effect of estrogen on the expression of long leptin receptor in rat adenohypophysis[J]. Jie Pou Xue Za Zhi, 2008, 31(6): 755-7. [陈红平, 张垒, 康路妹, 等. 雌激素对大鼠腺垂体中瘦素长型受体表达的影响[J]. 解剖学杂 志, 2008, 31(6): 755-7.] [8] Mentrikoski MJ, Shah AA, Hanley KZ, et al. Assessing endometrial hyperplasia and carcinoma treated with progestin therapy[J]. Am J Clin Pathol, 2012, 138(4): 524-34. [8] Mentrikoski MJ, Shah AA, Hanley KZ, et al. Assessing endometrial hyperplasia and carcinoma treated with progestin therapy[J]. Am J Clin Pathol, 2012, 138(4): 524-34. doi: 10.1309/AJCPM2TSDDF1MHBZ
[9] Lee LR, Teng PN, Nguyen H, et al. Progesterone enhances calcitriol antitumor activity by upregulating vitamin D receptor expression and promoting apoptosis in endometrial cancer cells[J]. Cancer Prev Res(Phila), 2013, 6(7): 731-43. [9] Lee LR, Teng PN, Nguyen H, et al. Progesterone enhances calcitriol antitumor activity by upregulating vitamin D receptor expression and promoting apoptosis in endometrial cancer cells[J]. Cancer Prev Res(Phila), 2013, 6(7): 731-43. doi: 10.1158/1940-6207.CAPR-12-0493
[10] Matsuoka S, Edwards MC, Bai C, et al. p57kip2, a structurally distinct member of the p21CIPI Cdk inhibitor family, is a candidate tumor suppressor gene[J]. Genes Dev, 1995, 9(6): 650-62. doi: 10.1101/gad.9.6.650
[10] Matsuoka S, Edwards MC, Bai C, et al. p57KIP2, a structurally distinct member of the p21CIPI Cdk inhibitor family, is a candidate tumor suppressor gene[J]. Genes Dev, l995, 9(6): 65 0-62. [11] Kavanagh E, Joseph B. The hallmarks of CDKN1C(p57, KIP2) in cancer[J]. Biochim Biophys Acta, 2011, 1816(1): 50-6. http://cn.bing.com/academic/profile?id=1997457074&encoded=0&v=paper_preview&mkt=zh-cn
[11] Kavanagh E, Joseph B. The hallmarks of CDKN1C( p57, KIP2) in cancer[J]. Biochim Biophys Acta, 2011, 1816(1): 50-6. [12] Mishra S, Lin CL, Huang TH, et al. MicroRNA-21 inhibits p57kip2 expression in prostate cancer[J]. Mol Cancer, 2014, 13: 212. doi: 10.1186/1476-4598-13-212
[12] Mishra S, Lin CL, Huang TH, et al. MicroRNA-21 inhibits p57Kip2 expression in prostate cancer[J]. Mol Cancer, 2014, 13: 212. [13] Li Y, Wang D, Zhang H, et al. P21-activated kinase 4 regulates the cyclin-dependent kinase inhibitor p57(kip2) in human breast cancer[J]. Anat Rec(Hoboken), 2013, 296(10): 1561-7. doi: 10.1002/ar.v296.10
[13] Li Y, Wang D, Zhang H, et al. P21-activated kinase 4 regulates the cyclin-dependent kinase inhibitor p57(kip2) in human breast cancer[J]. Anat Rec(Hoboken), 2013, 296(10): 1561-7. [14] Scaltriti M, Eichhorn PJ, Cortés J, et al. Cyclin E amplification/ overexpression is a mechanism of trastuzumab resistance in HER2+ breast cancer patients[J]. Proc Natl Acad Sci U S A, 2011, 10 8(9): 3761-6. [14] Scaltriti M, Eichhorn PJ, Cortés J, et al. Cyclin E amplification/overexpression is a mechanism of trastuzumab resistance in HER2+ breast cancer patients[J]. Proc Natl Acad Sci U S A, 2011, 108(9): 3761-6. doi: 10.1073/pnas.1014835108
[15] Zapiecki K, Manahan KJ, Miller GA, et al. Cyclin E is overexpressed by clear cell carcinomas of the endometrium and is a prognostic indicator of survival[J]. Eur J Gynaecol Oncol, 2015, 36 (2): 114-6. [15] Zapiecki K, Manahan KJ, Miller GA, et al. Cyclin E is overexpressed by clear cell carcinomas of the endometrium and is a prognostic indicator of survival[J]. Eur J Gynaecol Oncol, 2015, 36(2): 114-6. http://cn.bing.com/academic/profile?id=2340827538&encoded=0&v=paper_preview&mkt=zh-cn
[16] Zhou ZP, Xiao QB, Luo ZQ. Relationship between expression of p57kip2 and cyclin E proteins and clinicopathologic features in endometrial adenocarcinoma tissues[J]. Zhong Liu Fang Zhi Yan Jiu, 2013, 40(9): 860-3. [周正平, 肖庆邦, 罗祖强. 子宫内膜样腺 癌组织中p57kip2、cyclin E蛋白表达与临床病理特征的关系[J]. 肿瘤防治研究, 2013, 40(9): 860-3.] [16] 周正平, 肖庆邦, 罗祖强.子宫内膜样腺癌组织中p57kip2、cyclin E蛋白表达与临床病理特征的关系[J].肿瘤防治研究, 2013, 40(9): 860-3. http://www.zlfzyj.com/CN/abstract/abstract4362.shtml Zhou ZP, Xiao QB, Luo ZQ. Relationship between expression of p57kip2 and cyclin E proteins and clinicopathologic features in endometrial adenocarcinoma tissues[J]. Zhong Liu Fang Zhi Yan Jiu, 2013, 40(9): 860-3. http://www.zlfzyj.com/CN/abstract/abstract4362.shtml
[17] Darzynkiewicz Z, Zhao H, Zhang S, et al. Initiation and termination of DNA replication during S phase in relation to cyclins D1, E and A, p21WAF1, Cdt1 and the p12 subunit of DNA polymerase δ revealed in individual cells by cytometry[J]. Oncotarget, 2015, 6( 14): 11735-50. [17] Darzynkiewicz Z, Zhao H, Zhang S, et al. Initiation and termination of DNA replication during S phase in relation to cyclins D1, E and A, p21WAF1, Cdt1 and the p12 subunit of DNA polymeraseδrevealed in individual cells by cytometry[J]. Oncotarget, 2015, 6(14): 11735-50. doi: 10.18632/oncotarget
[18] Huang Y, Yoon MK, Otieno S, et al. The activity and stability of the intrinsically disordered Cip/Kip protein family are regulated by non-receptor tyrosine kinases[J]. J Mol Biol, 2015, 427(2): 371-86. doi: 10.1016/j.jmb.2014.11.011
[18] Huang Y, Yoon MK, Otieno S, et al. The activity and stability of the intrinsically disordered Cip/Kip protein family are regulated by non-receptor tyrosine kinases[J]. J Mol Biol, 2015, 427(2): 37 1-86.
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