Mechanism of circ0006152/miR-557/Wnt3a signal axis in the genesis and development of non-small cell lung cancer A549 cells
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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。 -
关键词:
- 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. -
Key words:
- 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
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
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
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
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
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±0.26a 0.57±0.05a CRL5800 10 2.64±0.32a 0.49±0.03a H226 10 2.69±0.31a 0.47±0.03a A549 10 3.83±0.45b 0.29±0.04b 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±5.36a 43.53±6.27a 58.29±9.16a 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±5.36a 42.53±6.27a 58.29±9.16a si-circ0006152+miR-557 inhibitors组 10 19.84±3.79a 21.79±4.69ab 110.79±12.84ab 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±0.07a 0.21±0.03a si-circ0006152+miR-557 inhibitors组 10 0.62±0.05ab 0.57±0.08ab 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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