circ0006152/miR-557/Wnt3a信号轴在非小细胞肺癌A549细胞发生发展中的机制

王近瑜; 李国平

doi:10.16766/j.cnki.issn.1674-4152.003416

circ0006152/miR-557/Wnt3a信号轴在非小细胞肺癌A549细胞发生发展中的机制

doi: 10.16766/j.cnki.issn.1674-4152.003416

王近瑜,
李国平^,

湖州市中心医院呼吸科，浙江湖州 313000

基金项目:

浙江省医药卫生科技计划项目 2019RC281

详细信息

通讯作者:
李国平，E-mail：sgit59@163.com

中图分类号: R734.2 R730.23
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- 文章访问数: 139
- HTML全文浏览量: 71
- PDF下载量: 6
- 被引次数: 0
出版历程
- 收稿日期: 2023-02-14
- 网络出版日期: 2024-05-27

Mechanism of circ0006152/miR-557/Wnt3a signal axis in the genesis and development of non-small cell lung cancer A549 cells

WANG Jinyu,
LI Guoping^,

Respiratory Department of Huzhou Central Hospital, Huzhou, Zhejiang 313000, China

摘要

摘要: 目的环状RNA(circRNA)与非小细胞肺癌发生发展有关，本研究旨在探讨circ0006152/miR-557/Wnt3a信号轴在非小细胞肺癌A549细胞发生发展中的机制。方法取对数生长期A549细胞进行分组和转染，采用随机数字法分为对照组(常规培育)、si-NC组(转染si-NC)、si-circ0006152组(转染si-circ0006152)、miR-NC组(转染miR-NC)、miR-557 mimics组(转染miR-557 mimics)和si-circ0006152+miR-557 inhibitors组(转染miR-557 inhibitors的同时进行si-circ0006152转染)，检测A549细胞增殖抑制率、凋亡、侵袭和Wnt3a、β-catenin蛋白表达。结果与si-NC组比较，si-circ0006152组的A549细胞增殖抑制率和凋亡率明显升高, 侵袭数量显著降低(P < 0.001);与miR-NC组比较，miR-557 mimics组的A549细胞增殖抑制率[(48.42±5.73)% vs. (4.27±0.19)%]、凋亡率[(45.38±5.96)% vs. (8.94±2.11)%]显著增加，细胞侵袭数量[(55.42±8.65)个vs. (167.42±19.53)个]明显降低(P＜0.001)；与si-circ0006152组比较，si-circ0006152+miR-557 inhibitors组中细胞增殖抑制率、凋亡率显著降低, 细胞侵袭数量明显增加(P < 0.05)，si-circ0006152+miR-557 inhibitors组中Wnt3a和β-catenin蛋白的表达显著升高(P＜0.05)。结论沉默circ000615可以通过调节miR-557/Wnt3a信号轴降低非小细胞肺癌细胞的恶性程度。
- 非小细胞肺癌 /
- Circ0006152 /
- miR-557 /
- Wnt3a /
- 机制
Abstract: Objective To investigate circular RNA is associated with the occurrence and development of non-small cell lung cancer. This study aims to investigate the mechanism of circ0006152/miR-557/Wnt3a signaling axis in the occurrence and development of non-small cell lung cancer A549 cells. Methods Take A549 cells in logarithmic growth period for grouping and transfection, and divide them into control group (cells are only routinely cultured), si-NC group (transfected with si-NC), si-circ0006152 group (transfected with si-circ0006152), miR-NC group (transfected with miR-NC), miR-557 mimics group (transfected with miR-557 mimics) and si-circ0006152+miR-557 inhibitors group (transfected with miR-557 inhibitors and si-circ0006152) by random number table method. Proliferation, apoptosis, migration invasion and Wnt3a, beta-catenin protein were detected. Results Compared with the si-NC group, the proliferation inhibition rate and apoptosis rate of A549 cells in the si-circ0006152 group were significantly increased, and the number of invasion was significantly decreased (P < 0.001); Compared with the miR-NC group, the proliferation inhibition rate of A549 cells [(48.42 ± 5.73)% vs.(4.27 ± 0.19)%] and apoptosis rate of A549 cells in miR-557 mimics group [(45.38±5.96)% vs. (8.94±2.11)%] were significantly increased. The number of cells invaded (55.42±8.65 vs. 167.42 ± 19.53) was significantly decreased (P < 0.001). The cell proliferation inhibition rate and apoptosis rate in the si-circ0006152+miR-557 inhibitors group were significantly decreased, and the number of cell invasion was significantly increased in the si-circ0006152+miR-557 inhibitors group (P < 0.05). The expressions of Wnt3a and β-catenin proteins in the inhibitors group were significantly increased (P < 0.05). Conclusion Silencing circ0006152 can reduce the malignancy of NSCLC A549 cells by regulating the miR-557/Wnt3a signaling axis.
- Non small cell lung cancer /
- Circ0006152 /
- miR-557 /
- Wnt3a /
- Mechanism

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图 1 各组A549细胞凋亡和侵袭能力

Figure 1. Apoptosis and invasion ability of A549 cells in each group

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图 2 各组A549细胞凋亡和侵袭能力

Figure 2. Apoptosis and invasion ability of A549 cells in each group

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图 3 circ0006152和miR-557核苷酸序列的结合位点

Figure 3. Binding sites between circ0006152 and miR-557

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图 4 各组A549细胞凋亡和侵袭能力

Figure 4. Apoptosis and invasion ability of A549 cells in each group

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图 5 各组Wnt3a和β-catenin蛋白的表达

Figure 5. Expression levels of Wnt3a and β-catenin in each group

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表 1 非小细胞肺癌细胞系中circ0006152和miR-557的表达水平比较(x±s)

Table 1. Comparison of the expression levels of circ0006152 and miR-557 in non-small cell lung cancer cell lines (x±s)

细胞系	n	circ0006152	miR-557
BEAS-2B	10	0.98±0.09	1.01±0.12
H292	10	2.03±0.26^a	0.57±0.05^a
CRL5800	10	2.64±0.32^a	0.49±0.03^a
H226	10	2.69±0.31^a	0.47±0.03^a
A549	10	3.83±0.45^b	0.29±0.04^b
F值		113.600	177.500
P值		＜0.001	＜0.001
注：与BEAS-2B比较，^aP＜0.05，^bP＜0.001。

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表 2 circ0006152对A549细胞增殖、凋亡和侵袭能力比较(x±s)

Table 2. Comparison of circ0006152 on the proliferation, apoptosis, and invasion of A549 cells (x±s)

组别	n	增殖抑制率(%)	凋亡率(%)	侵袭数量(个)
对照组	10	4.27±0.15	9.73±2.01	169.53±18.92
si-NC组	10	4.38±0.19	9.86±1.93	165.49±19.84
si-circ0006152组	10	47.57±5.36^a	43.53±6.27^a	58.29±9.16^a
F值		649.600	241.700	142.900
P值		＜0.001	＜0.001	＜0.001
注：与si-NC组比较，^aP＜0.001。

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表 3 miR-557对A549细胞增殖、凋亡和侵袭能力比较(x±s)

Table 3. Comparison of miR-557 on the proliferation, apoptosis and invasion of A549 cells (x±s)

组别	n	增殖抑制率(%)	凋亡率(%)	侵袭数量(个)
miR-NC组	10	4.27±0.19	8.94±2.11	167.42±19.53
miR-557 mimics组	10	48.42±5.73	45.38±5.96	55.42±8.65
t值		24.352	18.226	16.581
P值		＜0.001	＜0.001	＜0.001

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表 4 双荧光素酶活性检测结果(x±s)

Table 4. Results of dual-luciferase reporter assay (x±s)

组别	n	WT-circ0006152	MUT-circ0006152
miR-NC组	10	1.01±0.07	1.03±0.05
miR-557 mimics组	10	0.37±0.05	0.99±0.08
t值		23.527	0.098
P值		＜0.001	0.197

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表 5 各组A549细胞增殖、凋亡和侵袭能力比较(x±s)

Table 5. Comparison of proliferation, apoptosis and invasion of A549 cells in each group (x±s)

组别	n	增殖抑制率(%)	凋亡率(%)	侵袭数量(个)
si-NC组	10	4.38±0.19	9.86±1.93	165.49±19.84
si-circ0006152组	10	47.57±5.36^a	42.53±6.27^a	58.29±9.16^a
si-circ0006152+miR-557 inhibitors组	10	19.84±3.79^a	21.79±4.69^ab	110.79±12.84^ab
F值		333.100	126.100	134.200
P值		＜0.001	＜0.001	＜0.001
注：与si-NC组比较，^aP＜0.05；与si-circ0006152组比较，^bP＜0.05。

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表 6 各组Wnt3a信号轴相关蛋白表达比较(x±s)

Table 6. Comparison of protein expression related to the Wnt3a signaling pathway across all groups (x±s)

组别	n	Wnt3a	β-catenin
si-NC组	10	1.04±0.19	1.02±0.21
si-circ0006152组	10	0.27±0.07^a	0.21±0.03^a
si-circ0006152+miR-557 inhibitors组	10	0.62±0.05^ab	0.57±0.08^ab
F值		102.500	96.130
P值		＜0.001	＜0.001
注：与si-NC组比较，^aP＜0.05；与si-circ0006152组比较，^bP＜0.05。

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参考文献(17)

[1]	LI L, SUN D, LI X, et al. Identification of key circRNAs in non-small cell lung cancer[J]. Am J Med Sci, 2021, 361(1): 98-105. doi: 10.1016/j.amjms.2020.08.008
[2]	REN Y, SHE Y, TANG H, et al. Prognostic evaluation of the proposed residual tumor classification in a Chinese non-small cell lung cancer population[J]. J Surg Oncol, 2022, 125(6): 1061-1070. doi: 10.1002/jso.26810
[3]	ZHANG Q, KANG L K, LI X Y, et al. Bioinformatics analysis predicts hsa_circ_0026337/miR-197-3p as a potential oncogenic ceRNA network for non-small cell lung cancers[J]. Anticancer Agents Med Chem, 2022, 22(5): 874-886. doi: 10.2174/1871520621666210712090721
[4]	YANG Y, SUN K K, SHEN X J, et al. MiR-557 inhibits the proliferation and invasion of pancreatic cancer cells by targeting EGFR[J]. Int J Clin Exp Pathol, 2019, 12(4): 1333-1341.
[5]	BORA Y, TRIKALINOS N A, Vincent M, et al. Real-world use and survival outcomes of immune checkpoint inhibitors in older adults with non-small cell lung cancer[J]. Cancer, 2020, 126(5): 978-985. doi: 10.1002/cncr.32624
[6]	谢忠海, 李鸿伟, 臧金, 等. 非小细胞肺癌根治术患者术后临床特征调查及预后影响因素分析[J]. 中华全科医学, 2022, 20(11): 1860-1862. doi: 10.16766/j.cnki.issn.1674-4152.002720
[7]	LI M, LIU Y, JIANG X, et al. Inhibition of miR-144-3p exacerbates non-small cell lung cancer progression by targeting CEP55[J]. Acta Biochimit Biophys Sin, 2021, 53(10): 1398-1407. doi: 10.1093/abbs/gmab118
[8]	BALENDRAN P, PRICE M, TIWARI A, et al. Impact of PD1 and PDL1 immunotherapy on non-small cell lung cancer outcomes: a systematic review[J]. Thorax, 2022, 68(11): 394-424.
[9]	侯岩君, 田进海, 王立斌, 等. CircRNA hsa_circ_0044569在肺癌诊断中的价值[J]. 肿瘤防治研究, 2021, 48(12): 1061-1065. doi: 10.3971/j.issn.1000-8578.2021.21.0488
[10]	孔卓, 陈发坤, 夏彩霞, 等. Circ_0000212靶向miR-637调控肺癌H1299细胞的增殖、凋亡和放射敏感性[J]. 标记免疫分析与临床, 2022, 29(6): 1007-1014. https://www.cnki.com.cn/Article/CJFDTOTAL-BJMY202206022.htm
[11]	黎亮, 蔡仁中, 李高, 等. 非小细胞肺癌组织的环状RNAcirc_0080220水平和临床意义[J]. 临床肿瘤学杂志, 2021, 26(6): 537-540. https://www.cnki.com.cn/Article/CJFDTOTAL-LCZL202106008.htm
[12]	许俊, 朱承莹, 王德钧. circ_0015756通过调控miR-515-5p/HMGB3轴影响肺癌细胞增殖、凋亡和迁移[J]. 中国细胞生物学学报, 2021, 43(7): 1446-1454. https://www.cnki.com.cn/Article/CJFDTOTAL-XBZZ202107016.htm
[13]	陈绪林, 姜宇, 赵艳, 等. 原发性肝癌组织中miR-557的表达及其与预后的关系[J]. 国际消化病杂志, 2021, 41(4): 313-316. https://www.cnki.com.cn/Article/CJFDTOTAL-GWXH202104020.htm
[14]	ZHANG Y, ZHANG R, XI D, et al. EFNB2 acts as the target of miR-557 to facilitate cell proliferation, migration and invasion in pancreatic ductal adenocarcinoma by bioinformatics analysis and verification[J]. Am J Transl Res, 2018, 10(11): 3514-3528.
[15]	高志强, 李丁洋, 刘兆臣, 等. 环状RNA0007841靶向miR-557影响肝癌MHCC97H细胞增殖、迁移和侵袭的机制[J]. 中华实验外科杂志, 2021, 38(7): 1275-1279.
[16]	赛文莉, 姚敏, 郑文杰, 等. 癌组织Wnt3a异常表达及其靶向干预对肝癌的抑制作用[J]. 中华肝脏病杂志, 2019, 27(11): 866-871.
[17]	刘晶晶, 刘海娟, 王荣. Wnt3a对子宫内膜癌细胞转移的影响及其分子机制[J]. 山东医药, 2020, 60(29): 23-27.