核因子E2相关因子2基因沉默对人前列腺癌PC-3细胞增殖、侵袭和转移的影响

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摘要目的探讨核因子E2相关因子2(NRF2)基因沉默对人前列腺癌PC-3细胞增殖、侵袭和转移的影响及其机制。方法采用RT- PCR检测人前列腺上皮细胞株RWPE-1和前列腺癌细胞株PC-3中NRF2 mRNA的表达。采用脂质体转染法将NRF2 siRNA-1、NRF2 siRNA-2和NRF2 siRNA-3干扰序列转染至PC-3细胞后,Western blot和RT-PCR检测转染效果,并筛选出干扰效果最好的NRF2 siRNA序列。将体外培养的PC-3细胞分为Con组(未转染)、NC组(转染阴性对照)和NRF2 siRNA组(转染NRF2 siRNA-3),采用CCK-8法、平板克隆实验和Transwell实验分别检测各组细胞的增殖、克隆形成、侵袭和迁移能力,Western blot检测各组细胞中细胞核增殖相关抗原(Ki67)、细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶9(MMP-9)、N-钙黏附素(N-cadherin)蛋白的表达。结果与前列腺上皮细胞株RWPE-1相比,前列腺癌细胞株PC-3中NRF2 mRNA的表达水平明显升高(0.84±0.05 vs 0.22±0.03,P<0.05)。NRF2 siRNA-1、NRF2 siRNA-2和NRF2 siRNA-3干扰序列均能够明显下调PC-3细胞中NRF2蛋白(0.56±0.04,0.39±0.03,0.11±0.02 vs 0.76±0.07)和mRNA(0.80±0.05,0.52±0.03,0.26±0.03 vs 1.00±0.05)的表达,且NRF2 siRNA-3的干扰效果明显大于NRF2 siRNA-1和NRF2 siRNA-2。与Con组相比,NRF2 siRNA组细胞的增殖、克隆形成、侵袭和迁移能力[存活率:24 h (82.05±5.24)% vs (99.86±5.05)%,48 h (64.57±4.85)% vs (97.28±6.36)%,72 h (45.62±3.76)% vs (94.57±5.49)%;克隆形成率:(39.35±3.26)% vs (66.52±4.85)%;侵袭细胞数:42.00±5.50 vs 85.00±7.50;迁移细胞数:58.00±3.00 vs 118.50±8.00]均明显减弱,同时Ki67、CyclinD1、MMP-9和N-cadherin蛋白的表达(Ki67:0.25±0.03 vs 0.78±0.05;CyclinD1 0.41±0.03 vs 0.85±0.06;MMP-9:0.10±0.02 vs 0.89±0.07;N-cadherin:0.22±0.02 vs 0.49±0.04)均明显降低(P<0.05);而NC组与Con组间无统计学差异(P>0.05)。结论 NRF2基因沉默可抑制人前列腺癌PC-3细胞增殖、侵袭和转移,其分子机制可能与下调Ki67、CyclinD1、MMP-9和N-cadherin蛋白的表达有关。

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关键词 ：前列腺癌, NRF2基因, 细胞增殖, 细胞侵袭, 细胞迁移

Abstract：Objective To investigate the effect of NRF2 gene silencing on proliferation, invasion and metastasis of human prostate cancer PC-3 cells and the mechanism.Methods The expressions ofNRF2 mRNA in human prostate epithelial cell line RWPE-1 and prostate cancer cell line PC-3 were detected by RT-PCR.After the designed NRF2 siRNA-1, NRF2 siRNA-2 andNRF2 siRNA-3 interference sequences were transfected into PC-3 cells by lipofection, the bestNRF2 siRNA sequence was screened by Western blot and RT-PCR. PC-3 cells were divided into Con group (untransfected), NC group (transfected negative control) and NRF2siRNA group (transfected NRF2 siRNA-3). The proliferation, cloning, invasion and migration of each group were detected with CCK-8 method, plate cloning test and Transwell test,while the expressions of Ki67,CyclinD1, MMP-9 and N-cadherin proteins in each group were detected by Western blot.Results Compared with prostatic epithelial cell line RWPE-1, the expression of NRF2mRNA in prostate cancer cell line PC-3 was significantly increased (0.84±0.05 vs 0.22±0.03,P<0.05). The expressions of NRF2 protein(0.56±0.04,0.39±0.03,0.11±0.02 vs 0.76±0.07) and mRNA(0.80±0.05,0.52±0.03,0.26±0.03 vs 1.00±0.05) in PC-3 cells could be down-regulated by NRF2 siRNA-1, NRF2 siRNA-2 and NRF2 siRNA-3 interfering sequences,and the interference effect of NRF2 siRNA-3 was significantly greater than that of NRF2 siRNA-1 andNRF2 siRNA-2.Compared with Con group, the proliferation, cloning, invasion and migration abilities[survival rate:24 h (82.05±5.24)% vs (99.86±5.05)%,48 h (64.57±4.85)% vs(97.28±6.36)%,72 h (45.62±3.76)% vs(94.57±5.49)%;clone formation rate:(39.35±3.26)% vs(66.52±4.85)%;number of invasive cells:42.00±5.50 vs 85.00±7.50;number of migrating cells:58.00±3.00 vs 118.50±8.00]of NRF2 siRNA group were significantly weakened, while the expressions of Ki67, Cyclin D1, MMP-9 and N-cadherin (Ki67:0.25±0.03 vs 0.78±0.05;CyclinD1 0.41±0.03 vs 0.85±0.06;MMP-9:0.10±0.02 vs 0.89±0.07; N-cadherin:0.22±0.02 vs 0.49±0.04)were significantly decreased (P<0.05), but there was no significant difference between NC group and Con group (P>0.05).Conclusions NRF2gene silencing can inhibit the proliferation, invasion and metastasis of human prostate cancer PC-3 cells, and the molecular mechanism may be related to down-regulation of the expressions of Ki67, Cyclin D1, MMP-9 and N-cadherin proteins.

Key words： prostate cancer NRF2 gene cell proliferation cell invasion cell migration

收稿日期: 2019-03-15

ZTFLH:

R737.25

作者简介: 岳树香,本科学历,副主任中药师。

引用本文:

岳树香, 连玉菲, 刘广舒. 核因子E2相关因子2基因沉默对人前列腺癌PC-3细胞增殖、侵袭和转移的影响[J]. 武警医学, 2019, 30(7): 564-568. QIU Fuchun, LI Min, WANG Lu, WEI Wenpeng. Effect of NRF2 gene silencing on proliferation and metastasis of human prostate cancer PC-3 cells. Med. J. Chin. Peop. Armed Poli. Forc., 2019, 30(7): 564-568.

链接本文:

http://journal08.magtechjournal.com/Jwk_wjyx/CN/ 或 http://journal08.magtechjournal.com/Jwk_wjyx/CN/Y2019/V30/I7/564

[1]	周天域, 吕鑫, 陈雨桐,等.Nrf2的代谢调节作用与肿瘤的生长和增殖[J]. 中国细胞生物学学报, 2017,39(3):381-388.
[2]	Ahmed S M U, Luo L, Namani A, et al. Nrf2 signaling pathway: pivotal roles in inflammation[J]. Biochim Biophys Acta Mol Basis Dis, 2016, 1863(2):585-597.
[3]	Menegon S, Columbano A, Giordano S. The dual roles of NRF2 in cancer[J].Trends Mol Med, 2016, 22(7):578-593.
[4]	Kawasaki Y, Ishigami S, Arigami T, et al. Clinicopathological significance of nuclear factor (erythroid-2)-related factor 2 (Nrf2) expression in gastric cancer[J]. BMC Cancer,2015,15(1):1-8.
[5]	孙晓辉, 王彦, 徐畅,等. Nrf2基因的功能调节及其在肺癌中的作用机制研究进展[J]. 国际生物医学工程杂志, 2017,40(6):471-476.
[6]	Cho H Y, Kim K, Kim Y B, et al. Expression patterns of Nrf2 and keap1 in ovarian cancer cells and their prognostic role in disease recurrence and patient survival[J].Int J Gynecol Cancer, 2017, 27(3):412-419.
[7]	郝斌, 苗志刚, 袁媛,等. 前列腺癌组织中Nrf2、c-Myc蛋白表达变化及意义[J]. 山东医药, 2018,58(5):71-73.
[8]	Siegel R L, Miller K D, Jemal A. Cancer statistics, 2015[J].CA Cancer J Clin, 2015, 65(1): 5-29.
[9]	陈万青, 郑荣寿, 张思维,等. 2013年中国老年人群恶性肿瘤发病和死亡分析[J]. 中华肿瘤杂志, 2017, 39(1):60-66.
[10]	韩苏军, 张思维, 陈万青,等. 中国前列腺癌发病现状和流行趋势分析[J]. 临床肿瘤学杂志, 2013, 18(4):330-334.
[11]	Oudin M J, Weaver V M. Physical and chemical gradients in the tumor microenvironment regulate tumor cell invasion, migration, and metastasis[J]. Cold Spring HarbSymp Quant Biol, 2016, 81(1):189-205.
[12]	郑文键, 张协军, 黄国栋. Nrf2 信号通路在各系统肿瘤中的研究进展[J]. 中华临床医师杂志, 2014, 8(19): 3520-3524.
[13]	Kitano Y, Baba Y, Nakagawa S, et al. Nrf2 promotes oesophageal cancer cell proliferation via metabolic reprogramming and detoxification of reactive oxygen species[J]. J Pathol, 2018, 244(3): 346-357.
[14]	张钧, 孔德友, 张舰,等. RNA干扰Nrf2表达对食管癌生物学行为的影响[J]. 临床肿瘤学杂志, 2014,19(12):1057-1061.
[15]	Zhang C, Wang H J, Bao Q C, et al. NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction[J]. Oncotarget, 2016, 7(45):73593.
[16]	Duong H Q, You K S, Oh S, et al. Silencing of NRF2 reduces the expression of ALDH1A1 and ALDH3A1 and sensitizes to 5-FU in pancreatic cancer cells[J]. Antioxidants, 2017, 6(3):52-61.
[17]	Samatiwat P, Prawan A, Senggunprai L, et al. Nrf2 inhibition sensitizes cholangiocarcinoma cells to cytotoxic and antiproliferative activities of chemotherapeutic agents[J]. Tumour Biol, 2016, 37(8):11495-11507.
[18]	Green W J, Ball G, Hulman G, et al. KI67 and DLX2 predict increased risk of metastasis formation in prostate cancer-a targeted molecular approach[J]. Br J Cancer, 2016, 115(2):236-242.
[19]	赵明君, 王莹, 李巍伟. 前列腺癌组织内 PTTG1 基因表达量的变化及其与增殖, 侵袭基因的相关性研究[J]. 临床和实验医学杂志, 2018, 17(7): 710-713.