Nitrogen permease regulator like-2 regulates sensitivity of castration-resistant prostate cancer to olaparib

Xin CHEN; Wei1 TANG; Zhixiong CHEN; Bin ZHENG; Tao XIONG

doi:10.16016/j.1000-5404.201810053

您当前的位置：

首页 >

文章列表页 >

Nitrogen permease regulator like-2 regulates sensitivity of castration-resistant prostate cancer to olaparib

更新时间：2026-02-12

- Nitrogen permease regulator like-2 regulates sensitivity of castration-resistant prostate cancer to olaparib
- Journal of Third Military Medical University Vol. 41, Issue 3, Pages: 227-235(2019)
- 作者机构：
  
  1..400016 重庆, 重庆医科大学附属第一医院泌尿外科Department of Urology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016
  2..402460 重庆, 重庆市荣昌区人民医院泌尿外科Department of Urology, Rongchang District People's Hospital, Chongqing, 402460, China
- 作者简介：
- 基金信息：
  
  the General Program of Chongqing Municipal Health and Family Planning Commission(2016MSXM182)
- DOI：10.16016/j.1000-5404.201810053
  CLC： R73362;R737.25;R979.1
- Received：11 October 2018，
  
  Published：15 February 2019
- 稿件说明：
移动端阅览
Xin CHEN, Wei1 TANG, Zhixiong CHEN, et al. Nitrogen permease regulator like-2 regulates sensitivity of castration-resistant prostate cancer to olaparib[J]. Journal of Third Military Medical University, 2019, 41(3): 227-235.
DOI：

Xin CHEN, Wei1 TANG, Zhixiong CHEN, et al. Nitrogen permease regulator like-2 regulates sensitivity of castration-resistant prostate cancer to olaparib[J]. Journal of Third Military Medical University, 2019, 41(3): 227-235. DOI： 10.16016/j.1000-5404.201810053.

摘要

目的

探讨NPRL2在改善去势抵抗型前列腺癌(castration-resistant prostate cancer，CRPC)对Olaparib敏感性的作用。

方法

收集良性前列腺增生（benign prostatic hyperplasia，BPH）、普通前列腺癌(prostate cancer，PCa)和CRPC患者手术标本进行NPRL2的免疫组化染色，观察NPRL2在上述组织中的表达差异。Western blot检测正常前列腺上皮细胞RWPE-1、前列腺癌细胞LNCaP和耐恩杂鲁胺的去势抵抗型前列腺癌细胞LNPER中NPRL2蛋白的表达情况。通过慢病毒转染到CRPC细胞建立NPRL2的过表达和干扰，实时定量荧光PCR(qPCR)和Western blot验证NPRL2过表达和干扰效率

蛋白免疫印迹检测ATM蛋白表达。CCK-8法检测细胞增殖和细胞毒性，流式细胞术检测细胞凋亡，Transwell小室侵袭和迁移实验检测细胞侵袭转移。

结果

免疫组化结果显示NPRL2在CRPC组织中呈现高表达。Western blot结果显示NPRL2在RWPE-1细胞中低表达，在LNCaP和LNPER细胞中高表达，并成功构建过表达和干扰NPRL2的稳定株，qPCR及Western blot结果显示慢病毒转染能明显增高和降低NPRL2的mRNA和蛋白水平(

0.01)。CCK-8、流式细胞术和Transwell实验结果显示，与阴性对照比较，沉默NPRL2联合Olaparib能明显抑制细胞生长、阻止细胞侵袭转移和促进细胞凋亡(

0.01)。NPRL2与ATM表达呈正相关，NPRL2过表达联合Olaparib显著增加ATM表达。

结论

沉默NPRL2可增加CRPC细胞对Olaparib的敏感性。

Abstract

Objective

To investigate the effect of nitrogen permease regulator like-2 (NPRL2) on the sensitivity of castration-resistant prostate cancer (CRPC) to olaparib.

Methods

Immunohistochemical staining of NPRL2 was performed in surgical specimens from the patients with benign prostatic hyperplasia (BPH)

common prostate cancer (PCa) and CRPC. The expression of NPRL2 in the above tissues was observed and compared. Western blot assay was used to detect the protein level of NPRL2 in normal prostate epithelial cell line RWPE-1

prostate cancer cell line LNCaP and enzaliline-resistant castrated prostate cancer cell line LNPER. Overexpression and interference of NPRL2 were established by transfection of lentivirus into CRPC cells. The overexpression and interference efficiencies of NPRL2 were verified by real-time fluorescent PCR and Western blotting. ATM protein expression was detected by Western blotting. Cell proliferation and cytotoxicity were detected by CCK-8 assay

cell apoptosis was detected by flow cytometry

and invasion and migration were used with Transwell chamber test.

Results

Immunohistochemical assay showed that NPRL2 was highly expressed in the CRPC tissues. Western blotting showed that NPRL2 was down-regulated in RWPE-1 cells but highly expressed in LNCaP and LNPER cells. After successfully constructed a stable strain that overexpressed and interfered with NPRL2

qPCR and Western blotting showed that lentiviral transfection significantly increased and decreased the expression of NPRL2 at mRNA and protein levels

respectively(

0.01). And the results of CCK8 assay

flow cytometry and Transwell test indicated that silencing NPRL2 combined with olaparib treatment significantly inhibited cell growth

prevented cell invasion and metastasis

and promoted cell apoptosis when compared with the negative control group (

0.01). NPRL2 was positively correlated with ATM expression

and NPRL2 overexpression combined with olaparib treatment significantly enhanced ATM expression.

Conclusion

NPRL2 interference increases the sensitivity of CRPC cells to olaparib.

关键词

Keywords

references

ABESHOUSE A, AHN J, AKBANI R, et al.The molecular taxonomy of primary prostate cancer[J].Cell, 2015, 163(4):1011-1025.DOI:10.1016/j.cell.2015.10.025.

NA R, YE D, QI J, et al.Prostate health index significantly reduced unnecessary prostate biopsies in patients with PSA 2-10 ng/mL and PSA >10 ng/mL:Results from a multicenter study in China[J].Prostate, 2017, 77(11):1221-1229.DOI:10.1002/pros.23382.

ROSS R W, XIE W, REGAN M M, et al.Efficacy of androgen deprivation therapy (ADT) in patients with advanced prostate cancer:association between Gleason score, prostate-specific antigen level, and prior ADT exposure with duration of ADT effect[J].Cancer, 2008, 112(6):1247-1253.DOI:10.1002/cncr.23304.

ITO Y, SADAR M D.Enzalutamide and blocking androgen receptor in advanced prostate cancer:lessons learnt from the history of drug development of antiandrogens[J].Res Rep Urol, 2018, 10:23-32.DOI:10.2147/RRU.S157116.

MOTTET N, BELLMUNT J, BOLLA M, et al.EAU guidelines on prostate cancer.Part Ⅱ:Treatment of advanced, relapsing, and castration-resistant prostate cancer[J].Eur Urol, 2011, 59(4):572-583.DOI:10.1016/j.eururo.2011.01.025.

LERMAN M I, MINNA J D.The 630-kb lung cancer homozygous deletion region on human chromosome 3p21.3:identification and evaluation of the resident candidate tumor suppressor genes.The International Lung Cancer Chromosome 3p21.3 Tumor Suppressor Gene Consortium[J].Cancer Res, 2000, 60(21):6116-6133.

LI J, WANG F, HARALDSON K, et al.Functional characterization of the candidate tumor suppressor gene NPRL2/G21 located in 3p21.3C[J].Cancer Res, 2004, 64(18):6438-6443.DOI:10.1158/0008-5472.can-03-3869.

FARMER H, MCCABE N, LORD C J, et al.Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy[J].Nature, 2005, 434(7035):917-921.DOI:10.1038/nature03445.

ROBINSON D, VAN ALLEN E M, WU Y M, et al.Integrative clinical genomics of advanced prostate cancer[J].Cell, 2015, 161(5):1215-1228.DOI:10.1016/j.cell.2015.05.001.

JAYACHANDRAN G, UEDA K, WANG B, et al.NPRL2 sensitizes human non-small cell lung cancer (NSCLC) cells to cisplatin treatment by regulating key components in the DNA repair pathway[J].PLoS ONE, 2010, 5(8):e11994.DOI:10.1371/journal.pone.0011994.

LIU S, LIU B.Overexpression of nitrogen permease regulator like-2(NPRL2) enhances sensitivity to irinotecan (CPT-11) in colon cancer cells by activating the DNA damage checkpoint pathway[J].Med Sci Monit, 2018, 24:1424-1433.DOI:10.12659/msm.909186.

PENG Y, DAI H, WANG E, et al.TUSC4 functions as a tumor suppressor by regulating BRCA1 stability[J].Cancer Res, 2015, 75(2):378-386.DOI:10.1158/0008-5472.can-14-2315.

LIU M N, LIU A Y, PEI F H, et al.Functional mechanism of the enhancement of 5-fluorouracil sensitivity by TUSC4 in colon cancer cells[J].Oncol Lett, 2015, 10(6):3682-3688.DOI:10.3892/ol.2015.3801.

HUANG N, CHENG S, MI X, et al.Downregulation of nitrogen permease regulator like-2 activates PDK1-AKT1 and contributes to the malignant growth of glioma cells[J].Mol Carcinog, 2016, 55(11):1613-1626.DOI:10.1002/mc.22413.

TANG Y, JIANG L, TANG W.Decreased expression of NPRL2 in renal cancer cells is associated with unfavourable pathological, proliferation and apoptotic features[J].Pathol Oncol Res, 2014, 20(4):829-837.DOI:10.1007/s12253-014-9761-2.

BAR-PELED L, CHANTRANUPONG L, CHERNIACK A D, et al.A Tumor suppressor complex with GAP activity for the Rag GTPases that signal amino acid sufficiency to mTORC1[J].Science, 2013, 340(6136):1100-1106.DOI:10.1126/science.1232044.

CHEN Z, LUO S, CHEN Y, et al.High expression of NPRL2 is linked to poor prognosis in patients with prostate cancer[J].Hum Pathol, 2018, 76:141-148.DOI:10.1016/j.humpath.2018.02.011.

MATEO J, CARREIRA S, SANDHU S, et al.DNA-Repair defects and olaparib in metastatic prostate cancer[J].N Engl J Med, 2015, 373(18):1697-1708.DOI:10.1056/NEJMoa1506859.

ZHANG D, TANG B, XIE X, et al.The interplay between DNA repair and autophagy in cancer therapy[J].Cancer Biol Ther, 2015, 16(7):1005-1013.DOI:10.1080/15384047.2015.1046022.

LI Y, YANG D Q.The ATM inhibitor KU-55933 suppresses cell proliferation and induces apoptosis by blocking Akt in cancer cells with overactivated Akt[J].Mol Cancer Ther, 2010, 9(1):113-125.DOI:10.1158/1535-7163.mct-08-1189.

REDDY V, WU M, CIAVATTONE N, et al.ATM inhibition potentiates death of androgen receptor-inactivated prostate cancer cells with telomere dysfunction[J].J Biol Chem, 2015, 290(42):25522-25533.DOI:10.1074/jbc.M115.671404.

WESTON V J, OLDREIVE C E, SKOWRONSKA A, et al.The PARP inhibitor olaparib induces significant killing of ATM-deficient lymphoid tumor cells in vitro and in vivo [J ] .Blood, 2010, 116(22):4578-4587.DOI:10.1182/blood-2010-01-265769.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Expression of Reg3g after brain trauma and its effect on early epilepsy in mice

Reversing multi-cellular resistance of colon carcinoma cells by suppressing focal adhesion kinase through FAK/Akt/NF-κB signaling pathway

Related Author

Tianjun ZHOU

Saiyu CHENG

陈玉英

向浏欣

潘凤

蒋金妍

杨黎

李建军

Related Institution

Center of Neurological Diseases, the Third Affiliated Hospital of Chongqing Medical University

⁰