LncRNA SNHG17通过miR-384靶向AEG1对非小细胞肺癌细胞生物学行为的调控作用

doi:10.13481/j.1671-587X.20230205

Abstract

Abstract:

Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 17 （lncRNA SNHG17） on the biological behavior of the non-small cell lung cancer （NSCLC） cells， and to clarify its mechanism. Methods The human NSCLC A549 cells and H1299 cells were cultured，and the cells were transfected with pcDNA3.1-NC， SNHG17 over-expression plasmid（pcDNA3.1-SNHG17）， si-NC， SNHG17 small interference RNA（si-SNHG17），mimics NC，microRNA-384 mimics（miR-384 mimics）， inhibitor NC， miR-384 inhibitor（miR-384 inhibitor）， pcDNA3.1-astrocyte elevated gene 1 （AEG1）， and si-AEG1， respectively. Real-time quantitative PCR （RT-qPCR） method was used to detect the expression levels of lncRNA SNHG17，miR-384， and AEG1 mRNA in the A549 cells， H1299 cells and cells in various groups after transfection； ENCORI and TargetScan Databases were used to predicte the targeted bindings between lncRNA SNHG17 and miR-384，miR-384 and AEG1 mRNA. The A549 cells were divided into si-NC group， si-SNHG17 group， inhibitor NC group， miR-384 inhibitor group， si-NC+inhibitor NC group， si-SNHG17+inhibitor NC group， and si-SNHG17+miR-384 inhibitor group， si-AEG1 group， inhibitor NC+si-NC group， miR-384 inhibitor+si-NC group and miR-384 inhibitor+si-AEG1 group.The H1299 cells were divided into pcDNA3.1-NC group， pcDNA3.1-SNHG17 group， mimics NC group， miR-384 mimics group， pcDNA3.1-NC+mimics NC group， pcDNA3.1-SNHG17+mimics NC group，pcDNA3.1-SNHG17+miR-384 mimics group， pcDNA3.1-AEG1 group， mimics NC+pcDNA3.1-NC group， miR-384 mimics+pcDNA3.1-NC group， and miR-384 mimics+pcDNA3.1-AEG1 group. CCK-8 method was used to detect the cell viabilities in various groups； Transwell method was used to detect the numbers of migration and invasion cells in various groups； Western blotting method was used to detect the expression levels of AEG1 protein in the cells in various groups. Results The expression level of lncRNA SNHG17 in the H1299 cells was significantly lower than that in the A549 cells （P<0.01）. In the A549 cells， compared with si-NC group， the expression level of SNHG17 in the cells in si-SNHG17 group was significantly decreased （P<0.01），the cell viability， number of migration cells， and number of invasion cells were significantly decreased （P<0.01）； in the H1299 cells， compared with pcDNA3.1-NC group， the expression level of SNHG17 in the cells in pcDNA3.1-SNHG17 group was significantly increased （P<0.01）， the cell viability，number of migration cells， and number of invasion cells were increased significantly （P<0.01）. In the A549 cells， compared with si-NC group， the expression level of miR-384 in the cells in si-SNHG17 group was significantly increased（P<0.01）， and the expression level of AEG1 mRNA was significantly decreased（P<0.01）； in the H1299 cells， compared with pcDNA3.1-NC group， the expression level of miR-384 in the cells in pcDNA3.1-SNHG17 group was significantly decreased（P<0.01）， and the expression level of AEG1 mRNA was significantly increased （P<0.01）. The ENCORI Database results predicted that there were two binding sites between SNHG17 and miR-384. The rescue assay results showed that in the A549 cells， compared with si-NC+inhibitor NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in si-SNHG17+inhibitor NC group were significantly decreased（P<0.01）； compared with si-SNHG17+inhibitor NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in si-SNHG17+miR-384 inhibitor group were significantly increased（P<0.01）. In the H1299 cells， compared with pcDNA3.1-NC+mimics NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in pcDNA3.1-SNHG17+mimics NC group were significantly increased （P<0.01）； compared with pcDNA3.1-SNHG17+mimics NC group， the expression level of AEG1 protein， cell viability， number of migration cells， and number of invasion cells in pcDNA3.1-SNHG17+miR-384 mimics group were significantly decreased （P<0.01）. In the A549 cells， compared with si-NC group， the expression level of AEG1 protein in the cells， cell viability， number of migration cells， and number of invasion cells in si-AEG1 group were significantly decreased （P<0.01）； in the H1299 cells， compared with pcDNA3.1-NC group， the expression level of AEG1 protein cell viability， number of migration cells， and number of invasion cells were significantly increased （P<0.01）. The ENCORI Database results predicted that there was one binding site between miR-384 and AEG1.In the A549 cells， compared with inhibitor NC+si-NC group， the cell viability， number of migration cells and number of invasion cells in miR-384 inhibitor+si-NC group were significantly increased （P<0.01）； compared with miR-384 inhibitor+si-NC group， the cell viability， number of migration cells and number of invasion cells in miR-384 inhibitor+si-AEG1 group were significantly decreased （P<0.01）. In the H1299 cells， compared with the mimics NC+pcDNA3.1-NC group， the cell viability， number of migration cells， and number of invasion cells in miR-384 mimics+pcDNA3.1-NC group were significantly decreased （P<0.01）； compared with miR-384 mimics+pcDNA3.1-NC group， the cell viability， number of migration cells， and number of invasion cells in miR-384 mimics+pcDNA3.1-AEG1 group were significantly increased （P<0.01）. Conclusion LncRNA SNHG17 regulates the proliferation， migration， and invasion of the NSCLC cells by targeting AEG1 through miR-384.

Key words: Long non-coding RNA, Small nucleolar RNA host gene 17, MicroRNA-384, Astrocyte elevated gene 1, Cancer，Non-small cell lung, Biological behavior

CLC Number:

R734.2

Yunpeng LIU,Zihao LIU,Boming KANG,Zhiguang YANG. Regulatory effect of lncRNA SNHG17 on biological behavior of non-small cell lung cancer cells by targeting AEG1 through miR-384[J].Journal of Jilin University(Medicine Edition), 2023, 49(2): 298-307.

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References 25

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Group	lncRNA SNHG17	Cell viability (η/%)	Number of migration cells	Number of invasion cells
si-NC (A549 cells)	1.03±0.24	95.32±5.45	384.00±12.00	191.00±5.03
si-SNHG17 (A549 cells)	0.21±0.02^*	45.68±10.22^*	289.00±6.08^*	111.00±3.06^*
pcDNA3.1-NC (H1299 cells)	1.02±0.19	96.56±3.89	499.00±13.53	147.00±6.11
pcDNA3.1-SNHG17 (H1299 cells)	6.35±1.42^△	136.23±10.93^△	716.00±11.27^△	271.00±7.21^△

Group	miR-384	AEG1 mRNA
si-NC(A549 cells)	1.01±0.10	1.03±0.24
si-SNHG17(A549 cells)	1.46±0.20^*	0.06±0.02^*
pcDNA3.1-NC(H1299 cells)	1.01±0.13	1.02±0.19
pcDNA3.1-SNHG17(H1299 cells)	0.64±0.14^△	2.34±0.27^△

Group	miR-384	Cell viability (η/%)	Number of migration cells	Number of invasion cells
Inhibitor NC(A549 cells)	1.02±0.21	97.16±4.22	417.00±11.14	160.00±7.00
miR-384 inhibitor(A549 cells)	0.02±0.01^*	145.69±22.24^*	489.00±16.64^*	243.00±3.79^*
Mimics NC(H1299 cells)	1.02±0.21	96.37±3.22	518.00±11.53	207.00±6.03
miR-384 mimics(H1299 cells)	4.68±1.00^△	65.67±6.40^△	406.00±17.67^△	116.00±8.00^△

Group	Cell viability (η/%)	Number of migration cells	Number ofinvasion cells	Expression level of AEG1 protein
si-NC+inhibitor NC(A549 cells)	98.07±4.84	364.00±18.15	195.00±9.85	1.00±0.03
si-SNHG17+inhibitor NC(A549 cells)	63.26±6.04^*	314.00±8.72^*	108.00±6.11^*	0.72±0.01^*
si-SNHG17+miR-384 inhibitor(A549 cells)	78.69±6.65^△	350.00±8.74^△	199.00±11.15^△	0.81±0.01^△
pcDNA3.1-NC+mimics NC(H1299 cells)	101.26±5.91	496.00±18.23	148.00±4.58	1.00±0.02
pcDNA3.1-SNHG17+mimics NC(H1299 cells)	138.61±9.05^#	674.00±6.93^#	271.00±9.07^#	2.26±0.04^#
pcDNA3.1-SNHG17+miR-384 mimics(H1299 cells)	108.64±7.87^○	582.00±14.53^○	221.00±3.79^○	1.56±0.02^○

Group	AEG1 mRNA	Cell viability (η/%)	Number of migration cells	Number of invasion cells
si-NC(A549 cells)	1.02±0.22	98.67±4.76	381.00±13.08	167.00±6.03
si-AEG1(A549 cells)	0.56±0.08^*	71.32±8.98^*	308.00±10.26^*	115.00±10.02^*
pcDNA3.1-NC(H1299 cells)	1.01±0.11	99.21±7.66	499.00±11.53	137.00±7.09
pcDNA3.1-AEG1(H1299 cells)	3.15±0.90^△	132.32±9.33^△	609.00±11.14^△	282.00±3.79^△

Regulatory effect of lncRNA SNHG17 on biological behavior of non-small cell lung cancer cells by targeting AEG1 through miR-384

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Group	Cell viability (η/%)	Number of migration cells	Number of invasion cells
Inhibitor NC+si-NC(A549 cells)	101.36±7.40	408.00±12.58	161.00±6.11
miR-384 inhibitor+si-NC(A549 cells)	145.37±19.97^*	481.00±8.66^*	256.00±9.17^*
miR-384 inhibitor+si-AEG1(A549 cells)	112.36±7.47^△	425.00±9.07^△	172.00±9.07^△
Mimics NC+pcDNA3.1-NC(H1299 cells)	98.63±8.83	520.00±8.14	162.00±6.03
miR-384 mimics+pcDNA3.1-NC(H1299 cells)	63.17±9.12^#	401.00±10.44^#	117.00±2.08^#
miR-384 mimics+pcDNA3.1-AEG1(H1299 cells)	81.06±5.75^○	589.00±6.24^○	142.00±8.71^○

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