CDCA8通过调控增殖促进前列腺癌发生的作用机制研究

doi:10.11958/20212744

天津医药 ›› 2022, Vol. 50 ›› Issue (8): 796-801.doi: 10.11958/20212744

CDCA8通过调控增殖促进前列腺癌发生的作用机制研究

任智星¹(), 杨光华²^,^Δ()

1太原开放大学（邮编030024）
2山西白求恩医院,山西医学科学院,同济山西医院,山西医科大学第三医院泌尿外科

收稿日期:2021-12-12 修回日期:2022-01-20 出版日期:2022-08-15 发布日期:2022-08-12
通讯作者: 杨光华 E-mail:155151427@qq.com;836413443@qq.com
作者简介:任智星（1983）,女,讲师,主要从事临床医学相关的教学工作。E-mail： 155151427@qq.com
基金资助:
山西省基础研究计划项目(20210302123490)

Study on the mechanism of CDCA8 promoting the occurrence of prostate cancer by regulating proliferation

REN Zhixing¹(), YANG Guanghua²^,^Δ()

1 The Open University of Taiyuan, Taiyuan 030024, China
2 Department of Urology, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Third Hospital of Shanxi Medical University

Received:2021-12-12 Revised:2022-01-20 Published:2022-08-15 Online:2022-08-12
Contact: YANG Guanghua E-mail:155151427@qq.com;836413443@qq.com

任智星, 杨光华. CDCA8通过调控增殖促进前列腺癌发生的作用机制研究[J]. 天津医药, 2022, 50(8): 796-801.

REN Zhixing, YANG Guanghua. Study on the mechanism of CDCA8 promoting the occurrence of prostate cancer by regulating proliferation[J]. Tianjin Medical Journal, 2022, 50(8): 796-801.

摘要/Abstract

摘要：

目的探讨细胞分裂周期相关蛋白8（CDCA8）在前列腺癌（PCa）中的表达及作用机制。方法通过生物信息学方法分析正常前列腺组织和PCa组织中CDCA8 mRNA水平差异。利用癌症基因组图谱（TCGA）数据库中RNA表达测序数据分析CDCA8表达相关的PCa患者无病生存期（DFS）。免疫组织化学方法检测根治性前列腺切除术后56例PCa组织和癌旁组织中CDCA8蛋白表达差异,并依据染色指数将PCa患者分为CDCA8蛋白高表达组（31例）和低表达组（25例）,分析患者临床特征与CDCA8表达水平的相关性。利用siRNA沉默PCa细胞CDCA8基因,实时荧光定量聚合酶链反应（qPCR）检测CDCA8 mRNA的表达水平,细胞克隆形成实验检测PCa细胞增殖能力,蛋白免疫印迹法检测CDCA8、Ki67、增殖细胞核抗原（PCNA）以及磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（AKT）信号通路蛋白表达的变化。结果 TCGA数据库中PCa组织中CDCA8 mRNA水平明显高于正常组织,CDCA8 mRNA高表达的患者预后更差（P<0.01）。PCa组织CDCA8的蛋白表达水平明显高于癌旁组织,且表达水平与患者总前列腺特异性抗原（tPSA）、Gleason评分、T分期和术后切缘情况呈正相关（P<0.01）。CDCA8蛋白高表达组tPSA≥10 μg/L、T3期、术后切缘阳性患者比例大于CDCA8蛋白低表达组（P<0.05）。沉默CDCA8基因后PCa细胞增殖能力减弱,Ki67、PCNA、p-PI3K、p-AKT蛋白表达水平均下调（P<0.01）。结论 CDCA8通过调控增殖促进PCa发生,其作用机制可能与PI3K/AKT信号通路有关。

关键词: 前列腺肿瘤, 细胞增殖, 增殖细胞核抗原, 细胞周期蛋白质类, 细胞分裂周期相关蛋白8, PI3K/AKT信号通路

Abstract:

Objective To explore the expression and mechanism of cell division cycle associated protein 8 (CDCA8) in the occurrence of prostate cancer (PCa). Methods The difference of CDCA8 mRNA expression levels between normal prostate tissue and PCa tissue was analyzed by bioinformatics method. The disease-free survival (DFS) associated with CDCA8 expression in PCa patients was analyzed using RNA sequencing data from cancer Genome Atlas (TCGA) database. Immunohistochemical method was used to detect the expression of CDCA8 in 56 pairs of PCa tissues and adjacent tissues after radical prostatectomy. Patients were divided into the CDCA8 high expression group (n=31) and the CDCA8 low expression group (n=25) according to staining results. The potential correlation between clinical characteristics and CDCA8 expression level was further analyzed. The CDCA8 gene was silenced by siRNA, and the expression level of CDCA8 mRNA was detected by qPCR. The proliferation of PCa cells was detected by clonal formation assay, and the changes of CDCA8, Ki67, proliferating cell nuclear antigen (PCNA) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway proteins were detected by Western blot assay. Results In TCGA database, CDCA8 mRNA level was significantly higher in PCa tissues than that in normal tissues, and patients with high expression of CDCA8 mRNA had a worse prognosis (P<0.01). The staining index of expression level of CDCA8 protein was significantly higher in PCa tissue than that in para-cancer tissue, and the expression level was positively correlated with total prostate-specific antigen (tPSA), Gleason score, T stage and surgical margin (P<0.01). The proportion of patients with tPSA≥10 μg/L, T3 stage and positive postoperative resection margin was significantly higher in the high CDCA8 expression group than that in the low CDCA8 expression group (P<0.05). After CDCA8 gene was silenced, the proliferation of PCa cells was decreased, and the protein expression levels of Ki67, PCNA, p-PI3K and p-AKT were down-regulated (P<0.01). Conclusion CDCA8 promotes the occurrence of prostate cancer by regulating proliferation, and its mechanism may be related to PI3K/AKT signaling pathway.

Key words: prostatic neoplasms, cell proliferation, proliferating cell nuclear antigen, cell cycle proteins, CDCA8, PI3K/AKT signaling pathway

中图分类号:

R737.25

图/表 12

图1 CDCA8 mRNA高表达组和低表达组DFS比较

Fig.1 Comparison of DFS between the CDCA8 mRNA high expression group and the CDCA8 mRNA low expression group

图2 PCa组织中CDCA8的表达（免疫组化,左×100,右×200）

Fig.2 The expression of CDCA8 in PCa tissues (immunohistochemical staining, left×100, right×200)

图3 前列腺癌旁组织中CDCA8的表达（免疫组化,左×100,右×200）

Fig.3 The expression of CDCA8 in paracancerous tissues (immunohistochemical staining, left×100, right×200)

表1 2组前列腺癌患者的临床特征和CDCA8组织表达情况比较

Tab.1 Clinicopathological characteristics and CDCA8 expression of PCa patients between the two groups

组别	n	CDCA8 蛋白	年龄（岁）	tPSA（μg/L）		Gleason评分		T分期		N分期		术后切缘
组别	n	CDCA8 蛋白	年龄（岁）	<10	≥10	<7	≥7	T1~T2	T3	N0	N1	阴性	阳性
低表达组	25	1.92±0.28	68.84±6.23	9（36.0）	16（64.0）	5（20.0）	20（80.0）	19（76.0）	6（24.0）	24（96.0）	1（4.0）	19（76.0）	6（24.0）
高表达组	31	3.48±0.51	68.58±6.26	3（9.7）	28（90.3）	1（3.2）	30（96.8）	15（48.4）	16（51.6）	24（77.4）	7（22.6）	9（29.0）	22（71.0）
t或χ²		14.653^＊＊	0.154	5.695^＊＊		2.506		4.424^＊		2.532		12.212^＊＊

表2 Si-CDCA8组和Si-NC组CDCA8 mRNA及蛋白表达水平比较（n=3,$\bar{x}±s$）

Tab.2 Comparison of CDCA8 mRNA and protein expression between the Si-CDCA8 group and the Si-NC group

组别	CDCA8 mRNA		CDCA8蛋白
组别	PC3	DU145	PC3	DU145
Si-NC组	1.12±0.06	1.00±0.05	1.20±0.01	1.05±0.01
Si-CDCA8组	0.15±0.01	0.15±0.01	0.56±0.01	0.30±0.01
t	27.129^＊＊	28.121^＊＊	74.929^＊＊	147.377^＊＊

图4 Si-CDCA8组和Si-NC组CDCA8蛋白表达水平 A：Si-NC组;B：Si-CDCA8组;图5~7同。

Fig.4 Expression levels of CDCA8 proteins in the Si-CDCA8 group and the Si-NC group

表3 Si-CDCA8组和Si-NC组细胞集落数比较（n=3,个/视野,$\bar{x}±s$）

Tab.3 Comparison of cell colony number between the Si-CDCA8 group and the Si-NC group

组别	PC3	DU145
Si-NC组	316.67±6.43	522.67±4.04
Si-CDCA8组	160.67±14.01	348.33±7.37
t	17.527^＊＊	35.920^＊＊

图5 Si-CDCA8组和Si-NC组细胞集落数

Fig.5 The number of cell colonies in the Si-CDCA8 group and the Si-NC group

表4 Si-CDCA8组和Si-NC组Ki67和PCNA表达水平比较（n=3,$\bar{x}±s$）

Tab.4 Comparison of Ki67 and PCNA expression levels between the Si- CDCA8 group and the Si-NC group

组别	PC3		DU145
组别	Ki67	PCNA	Ki67	PCNA
Si-NC组	1.27±0.03	1.36±0.05	1.24±0.02	1.48±0.04
Si-CDCA8组	0.51±0.01	0.64±0.02	0.53±0.01	0.89±0.11
t	48.566^＊＊	22.695^＊＊	45.987^＊＊	8.868^＊＊

图6 Si-CDCA8组和Si-NC组Ki67和PCNA表达水平

Fig.6 Expression levels of Ki67 and PCNA in the Si-CDCA8 group and the Si-NC group

表5 Si-CDCA8组和Si-NC组PI3K/AKT通路蛋白表达水平比较（n=3,$\bar{x}±s$）

Tab.5 Comparison of PI3K /Akt pathway protein expression between the Si-CDCA8 group and the Si-NC group

组别	PC3
组别	PI3K	p-PI3K	AKT	p-AKT
Si-NC组	1.001±0.007	0.787±0.016	0.890±0.002	1.156±0.002
Si-CDCA8组	0.856±0.007	0.278±0.006	0.871±0.008	0.432±0.002
t	24.630^＊＊	50.049^＊＊	4.154^＊	382.818^＊＊
组别	DU145
组别	PI3K	p-PI3K	AKT	p-AKT
Si-NC组	0.676±0.004	0.925±0.023	0.991±0.006	0.841±0.005
Si-CDCA8组	0.610±0.004	0.360±0.008	1.013±0.010	0.410±0.003
t	18.051^＊＊	40.935^＊＊	3.164^＊	129.892^＊＊

图7 Si-CDCA8组和Si-NC组PI3K/AKT通路蛋白表达水平

Fig.7 Expression levels of PI3K/AKT pathway protein in the Si-CDCA8 group and the Si-NC group

参考文献 21

[1]	SIEGEL R L, MILLER K D, FUCHS H E, et al. Cancer Statistics,2021[J]. CA Cancer J Clin, 2021, 71(1):7-33. doi: 10.3322/caac.21654.
[2]	SIEGEL R L, MILLER K D, JEMAL A, et al. Cancer statistics,2020[J]. CA Cancer J Clin, 2020, 70(1):7-30. doi: 10.3322/caac.21590.
[3]	WONG M C, GOGGINS W B, WANG H H, et al. Global incidence and mortality for prostate cancer:analysis of temporal patterns and trends in 36 countries[J]. Eur Urol, 2016, 70(5):862-874. doi: 10.1016/j.eururo.2016.05.043.
[4]	WANG X, WANG H, XU J, et al. Double-targeting CDCA8 and E2F1 inhibits the growth and migration of malignant glioma[J]. Cell Death Dis, 2021, 12(2):146. doi: 10.1038/s41419-021-03405-4.
[5]	JEON T, KO M J, SEO Y R, et al. Silencing CDCA8 suppresses hepatocellular carcinoma growth and stemness via restoration of ATF3 tumor suppressor and inactivation of AKT/β-Catenin signaling[J]. Cancers(Basel), 2021, 13(5):1055. doi: 10.3390/cancers13051055.
[6]	BU Y, SHI L, YU D, et al. CDCA8 is a key mediator of estrogen-stimulated cell proliferation in breast cancer cells[J]. Gene, 2019, 703:1-6. doi: 10.1016/j.gene.2019.04.006.
[7]	CHEN C, CHEN S, PANG L, et al. Analysis of the expression of genes and its prognostic significance in human lung carcinoma:A review of the literature databases[J]. Biomed Res Int, 2020, 2020:6412593. doi: 10.1155/2020/6412593.
[8]	黄健. 中国泌尿外科和男科疾病诊断治疗指南[M]. 北京: 科学出版社, 2020:95-96.
	HUANG J. Guidelines for diagnosis and treatment of urology and andrology in China[M]. Beijing: Science Press, 2020:95-96.
[9]	YAN H, LI Z, SHEN Q, et al. Aberrant expression of cell cycle and material metabolism related genes contributes to hepatocellular carcinoma occurrence[J]. Pathol Res Pract, 2017, 213(4):316-321. doi: 10.1016/j.prp.2017.01.019.
[10]	HAASE J, BONNER M K, HALAS H, et al. Distinct roles of the chromosomal passenger complex in the detection of and response to errors in kinetochore-microtubule attachment[J]. Dev Cell, 2017, 42(6):640-654.e5. doi: 10.1016/j.devcel.2017.08.022.
[11]	ABAD M A, RUPPERT J G, BUZUK L, et al. Borealin-nucleosome interaction secures chromosome association of the chromosomal passenger complex[J]. J Cell Biol, 2019, 218(12):3912-3925. doi: 10.1083/jcb.201905040.
[12]	CI C, TANG B, LYU D, et al. Overexpression of CDCA8 promotes the malignant progression of cutaneous melanoma and leads to poor prognosis[J]. Int J Mol Med, 2019, 43:404-412. doi: 10.3892/ijmm.2018.3985.
[13]	LI Q, LIANG J, CHEN B. Identification of CDCA8,DSN1 and BIRC5 in regulating cell cycle and apoptosis in osteosarcoma using bioinformatics and cell biology[J]. Technol Cancer Res Treat, 2020, 19:1533033820965605. doi: 10.1177/1533033820965605.
[14]	CHANG J L, CHEN T H, WANG C F, et al. Borealin/Dasra B is a cell cycle-regulated chromosomal passenger protein and its nuclear accumulation is linked to poor prognosis for human gastric cancer[J]. Exp Cell Res, 2006, 312(7):962-973. doi: 10.1016/j.yexcr.2005.12.015.
[15]	HAYAMA S, DAIGO Y, YAMABUKI T, et al. Phosphorylation and activation of cell division cycle associated 8 by aurora kinase B plays a significant role in human lung carcinogenesis[J]. Cancer Res, 2007, 67(9):4113-4122. doi: 10.1158/0008-5472.CAN-06-4705.
[16]	BI Y, CHEN S, JIANG J, et al. CDCA8 expression and its clinical relevance in patients with bladder cancer[J]. Medicine(Baltimore), 2018, 97(34):e11899. doi: 10.1097/MD.0000000000011899.
[17]	CHEN X, SUN J, WANG Y. Expressions of CD44,PCNA and MRP1 in lung cancer tissues and their effects on proliferation and invasion abilities of lung cancer cell line 95D[J]. J BUON, 2021, 26(1):72-78.
[18]	MENON S S, GURUVAYOORAPPAN C, SAKTHIVEL K M, et al. Ki-67 protein as a tumour proliferation marker[J]. Clin Chim Acta, 2019, 491:39-45. doi: 10.1016/j.cca.2019.01.011.
[19]	KREIS N N, LOUWEN F, YUAN J. The Multifaceted p21(Cip1/Waf1/CDKN1A)in cell differentiation,migration and cancer therapy[J]. Cancers(Basel), 2019, 11(9):1220. doi: 10.3390/cancers11091220.
[20]	MU M, NIU W, ZHANG X, et al. LncRNA BCYRN1 inhibits glioma tumorigenesis by competitively binding with miR-619-5p to regulate CUEDC2 expression and the PTEN/AKT/p21 pathway[J]. Oncogene, 2020, 39(45):6879-6892. doi: 10.1038/s41388-020-01466-x.
[21]	CUI X H, CHEN T H, LI R Z, et al. Cell division cycle associated 8:A novel diagnostic and prognostic biomarker for hepatocellular carcinoma[J]. J Cell Mol Med, 2021, 25(24):11097-11112. doi: 10.1111/jcmm.17032.

CDCA8通过调控增殖促进前列腺癌发生的作用机制研究

Study on the mechanism of CDCA8 promoting the occurrence of prostate cancer by regulating proliferation

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