长链非编码RNA ARAP1-AS1在肾透明细胞癌中的表达及作用研究

doi:10.19401/j.cnki.1007-3639.2022.01.004

中国癌症杂志 ›› 2022, Vol. 32 ›› Issue (1): 34-40.doi: 10.19401/j.cnki.1007-3639.2022.01.004

长链非编码RNA ARAP1-AS1在肾透明细胞癌中的表达及作用研究

雷坤阳¹, 谢文杰¹(), 孙庭¹, 刘贻富¹, 王旭²

1. 南昌大学第一附属医院泌尿外科,江西南昌 330006
2. 南昌大学第一附属医院病理科,江西南昌 330006

收稿日期:2021-10-07 修回日期:2021-11-25 出版日期:2022-01-30 发布日期:2022-01-25
通信作者: 谢文杰 E-mail:doctorxiewenjie@163.com
基金资助:
江西省卫生健康委科技计划项目(202210397)

Expression and effect of long non-coding RNA ARAP1-AS1 in clear cell renal cell carcinoma

LEI Kunyang¹, XIE Wenjie¹(), SUN Ting¹, LIU Yifu¹, WANG Xu²

1. Department of Urology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China
2. Department of Pathology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China

Received:2021-10-07 Revised:2021-11-25 Published:2022-01-30 Online:2022-01-25
Contact: XIE Wenjie E-mail:doctorxiewenjie@163.com

摘要/Abstract

摘要：

背景与目的：长链非编码RNA（long non-coding RNA,lncRNA）ARAP1-AS1在多种肿瘤中异常表达,但其在肾透明细胞癌（clear cell renal cell carcinoma,ccRCC）中的作用尚不清楚。探讨ARAP1-AS1在ccRCC中的生物学作用。方法：通过GEPIA数据库分析ARAP1-AS1在ccRCC组织中的表达及其与临床病理学特征及患者生存率的关系。采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR）检测ccRCC组织及邻近的非肿瘤组织中ARAP1-AS1的表达水平。将患者分为ARAP1-AS1高表达组和低表达组,分析ARAP1-AS1的表达水平与患者临床病理学特征之间的关系,并进行生存分析。通过细胞计数试剂盒-8（cell counting kit-8,CCK-8）实验、transwell迁移实验及侵袭实验检测ARAP1-AS1对ccRCC细胞体外增殖、迁移及侵袭能力的影响。采用蛋白质印迹法（Western blot）检测Wnt/β-catenin信号通路相关蛋白表达变化。采用BALB/c裸小鼠移植瘤模型分析ARAP1-AS1对ccRCC细胞体内成瘤能力的影响。结果：GEPIA数据库分析结果显示,ARAP1-AS1在ccRCC中高表达,且与患者肿瘤高分期及较差的生存率相关（P均<0.05）。RTFQ-PCR显示,ARAP1-AS1在ccRCC组织及细胞系中高表达,ARAP1-AS1的高表达与肿瘤大小和分期相关（P均<0.05）。ARAP1-AS1高表达患者的总生存率较差（P<0.05）。沉默ARAP1-AS1的表达可以抑制ccRCC细胞增殖、迁移和侵袭（P均<0.05）。沉默ARAP1-AS1可以降低Wnt/β-catenin信号通路相关蛋白的表达水平（P均<0.05）。沉默ARAP1-AS1可使ccRCC细胞体内成瘤能力减弱,并使Ki-67增殖指数降低。结论：ARAP1-AS1可通过激活Wnt/β-catenin信号通路促进ccRCC的进展。

关键词: 肾透明细胞癌, ARAP1-AS1, Wnt/β-catenin

Abstract:

Background and purpose: Long non-coding RNA (lncRNA) ARAP1-AS1 is abnormally expressed in a variety of tumors, but its role in clear cell renal cell carcinoma (ccRCC) is unknown. This study aimed to explore the biological role of ARAP1-AS1 in ccRCC. Methods: The expression level of ARAP1-AS1 in ccRCC tissues and its relationship to patient’s clinicopathological characteristics and survival rate were analyzed using the GEPIA database. The expression level of ARAP1-AS1 was measured in ccRCC and adjacent non-tumor tissues by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR). Patients were divided into ARAP1-AS1 high and low expression groups, the relationship between ARAP1-AS1 expression level and patient’s clinicopathological characteristics was analyzed, and survival analysis was performed. The effect of ARAP1-AS1 in vitro proliferation, migration and invasion ability of ccRCC cells was determined by cell counting kit-8 (CCK-8) assay, transwell migration and invasion assay. Changes in the expressions of Wnt/β-catenin signaling pathway-related proteins were detected by Western blot assay. The effect of ARAP1-AS1 on tumorigenic capacity in ccRCC cells in vivo was verified by tumor xenografts in nude mice. Results: The analysis of the GEPIA database showed that ARAP1-AS1 was highly expressed in ccRCC and associated with advanced tumor stage as well as poor survival in patients (all P<0.05). The results of RTFQ-PCR showed that ARAP1-AS1 was highly expressed in ccRCC tissues and cell lines, and high expression correlated with tumor size and stage (all P<0.05). The overall survival rate was poor in patients with high ARAP1-AS1 expression (P<0.05). Knockdown of ARAP1-AS1 expression inhibited the proliferation, migration and invasion of ccRCC cells (all P<0.05). Silencing of ARAP1-AS1 reduced the expression levels of the proteins involved in the Wnt/β-catenin signaling pathway (all P<0.05). Silencing of ARAP1-AS1 attenuated tumorigenic capacity in ccRCC cells and reduced Ki-67 proliferation index (P<0.05). Conclusion: ARAP1-AS1 promotes ccRCC progression through activation of the Wnt/β-catenin signaling pathway.

Key words: Clear cell renal cell carcinoma, ARAP1-AS1, Wnt/β-catenin

中图分类号:

R737.11

雷坤阳, 谢文杰, 孙庭, 刘贻富, 王旭. 长链非编码RNA ARAP1-AS1在肾透明细胞癌中的表达及作用研究[J]. 中国癌症杂志, 2022, 32(1): 34-40.

LEI Kunyang, XIE Wenjie, SUN Ting, LIU Yifu, WANG Xu. Expression and effect of long non-coding RNA ARAP1-AS1 in clear cell renal cell carcinoma[J]. China Oncology, 2022, 32(1): 34-40.

图/表 6

图1

图2

表1

图3

图4

图5

参考文献 17

[1]	HSIEH J J, PURDUE M P, SIGNORETTI S, et al. Renal cell carcinoma[J]. Nat Rev Dis Primers, 2017, 3: 17009. doi: 10.1038/nrdp.2017.9
[2]	STATELLO L, GUO C J, CHEN L L, et al. Gene regulation by long non-coding RNAs and its biological functions[J]. Nat Rev Mol Cell Biol, 2021, 22(2): 96-118. doi: 10.1038/s41580-020-00315-9
[3]	LUO H C, ZHU G Q, XU J F, et al. HOTTIP lncRNA promotes hematopoietic stem cell self-renewal leading to AML-like disease in mice[J]. Cancer Cell, 2019, 36(6): 645-659. e8. doi: 10.1016/j.ccell.2019.10.011
[4]	KIM J, PIAO H L, KIM B J, et al. Long non-coding RNA MALAT1 suppresses breast cancer metastasis[J]. Nat Genet, 2018, 50(12): 1705-1715. doi: 10.1038/s41588-018-0252-3
[5]	LI M Z, YIN B D, CHEN M L, et al. Downregulation of the lncRNA ASB16-AS1 decreases LARP1 expression and promotes clear cell renal cell carcinoma progression via miR-185-5p/miR-214-3p[J]. Front Oncol, 2020, 10: 617105. doi: 10.3389/fonc.2020.617105
[6]	ZHU K, MIAO C K, TIAN Y, et al. lncRNA MIR4435-2HG promoted clear cell renal cell carcinoma malignant progression via miR-513a-5p/KLF6 axis[J]. J Cell Mol Med, 2020, 24(17): 10013-10026. doi: 10.1111/jcmm.v24.17
[7]	TENG J F, AI X, JIA Z M, et al. Long non-coding RNA ARAP1-AS1 promotes the progression of bladder cancer by regulating miR-4735-3p/NOTCH2 axis[J]. Cancer Biol Ther, 2019, 20(4): 552-561. doi: 10.1080/15384047.2018.1538613
[8]	LU C, WANG X H, ZHAO X W, et al. Long non-coding RNA ARAP1-AS1 accelerates cell proliferation and migration in breast cancer through miR-2110/HDAC2/PLIN1 axis[J]. Biosci Rep, 2020, 40(4): BSR20191764.
[9]	YE Y Q, GU B B, WANG Y, et al. YY1-induced upregulation of long non-coding RNA ARAP1-AS1 promotes cell migration and invasion in colorectal cancer through the wnt/β-catenin signaling pathway[J]. Cancer Biother Radiopharm, 2019, 34(8): 519-528. doi: 10.1089/cbr.2018.2745
[10]	CHEN Y Y, LI Z J, CHEN X G, et al. Long non-coding RNAs: from disease code to drug role[J]. Acta Pharm Sin B, 2021, 11(2): 340-354. doi: 10.1016/j.apsb.2020.10.001
[11]	MING H, LI B W, ZHOU L, et al. Long non-coding RNAs and cancer metastasis: molecular basis and therapeutic implications[J]. Biochim Biophys Acta Rev Cancer, 2021, 1875(2): 188519. doi: 10.1016/j.bbcan.2021.188519
[12]	ZHANG Y, WU D, WANG D. Long non-coding RNA ARAP1-AS1 promotes tumorigenesis and metastasis through facilitating proto-oncogene c-Myc translation via dissociating PSF/PTB dimer in cervical cancer[J]. Cancer Med, 2020, 9(5): 1855-1866. doi: 10.1002/cam4.v9.5
[13]	JIANG Y H, ZHU X Y, GUO Z Y, et al. Increased long non-coding RNA ARAP1-AS1 expression and its prognostic significance in human gastric cancer: a preliminary study[J]. Eur Rev Med Pharmacol Sci, 2020, 24(4): 1815-1820.
[14]	TAO X L, ZHANG Y, LI J P, et al. Low expression of long non-coding RNA ARAP1-AS1 can inhibit lung cancer proliferation by inducing G₀/G₁ cell cycle organization[J]. J Thorac Dis, 2020, 12(12): 7326-7336. doi: 10.21037/jtd
[15]	TREJO-SOLIS C, ESCAMILLA-RAMIREZ A, JIMENEZ-FARFAN D, et al. Crosstalk of the wnt/β-catenin signaling pathway in the induction of apoptosis on cancer cells[J]. Pharmaceuticals (Basel), 2021, 14(9): 871. doi: 10.3390/ph14090871
[16]	XU X L, ZHANG N W, GAO R X, et al. Upregulation of SDHA inhibited proliferation, migration, and invasion of clear cell renal cell carcinoma cells via inactivation of the Wnt/β-catenin pathway[J]. J Recept Signal Transduct Res, 2021: 1-13.
[17]	ZHU L, LIU X L, FU Z Q, et al. LINC00675 suppresses proliferative, migration and invasion of clear cell renal cell carcinoma via the Wnt/β-catenin pathway[J]. Eur Rev Med Pharmacol Sci, 2020, 24(5): 2313-2320.

Clinicopathological feature	ARAP1-AS1 expression		P value
Clinicopathological feature	High (n=32)	Low (n=33)	P value
Age/year			0.378
≤60	13	17
>60	19	16
Gender			0.174
Male	15	21
Female	17	12
Tumor size D/cm			0.032
<4	10	19
≥4	22	14
Tumor stage			0.035
Ⅰ and Ⅱ	12	21
Ⅲ and Ⅳ	20	12
Lymph node metastasis			0.389
Positive	15	12
Negative	17	21
Distant metastasis			0.265
Positive	14	19
Negative	18	14

长链非编码RNA ARAP1-AS1在肾透明细胞癌中的表达及作用研究

Expression and effect of long non-coding RNA ARAP1-AS1 in clear cell renal cell carcinoma

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 17

相关文章 3

编辑推荐

Metrics

本文评价

[1]	王　佳, 艾中中, 周　菁 . 下调SLC6A3对肾透明细胞癌细胞SNU-349增殖、迁移的影响[J]. 中国癌症杂志, 2021, 31(1): 20-26.
[2]	童　颖, 余怡雯, 谢素红, 王砚春, 卢仁泉, 郭　林. 沉默信息调节因子过表达或谷氨酰胺剥夺对肾透明细胞癌细胞凋亡、增殖的影响[J]. 中国癌症杂志, 2020, 30(6): 435-440.
[3]	孙小亮,鲁　瑶,杨　猛. 肾透明细胞癌术后双侧甲状腺转移1例[J]. 中国癌症杂志, 2017, 27(10): 839-840.