Effects of lupeol on the biological behavior of non-small cell lung cancer NCI-H1299 cells by regulating miR-100-5p

doi:10.11958/20220899

Abstract

Abstract:

Objective To investigate the effect of lupeol on the proliferation, migration and invasion of non-small cell lung cancer (NSCLC) NCI-H1299 cells by regulating miR-100-5p. Methods NCI-H1299 cells cultured in vitro were divided into the normal control (NC) group (untreated), the low-dose group (12.5 μmol/L lupeol), the medium-dose group (25 μmol/L lupeol), the high-dose group (50 μmol/L lupeol), the miR-NC group (transfected with miR-NC), the miR-100-5p group (transfected with miR-100-5p), the anti-miR-NC+high-dose group (the cells were treated with 50 μmol/L lupeol after transfected with anti-miR-NC), and the anti-miR-100-5p+high-dose group (the cells were treated with 50 μmol/L lupeol after transfected with anti-miR-100-5p). CCK-8 and colony formation assays were performed to evaluate the proliferation of NCI-H1299 cells. Transwell method was applied to detect the migration and invasion of NCI-H1299 cells. RT-qPCR was used to detect the expression of miR-100-5p. Western blot assay was used to detect the expression of CyclinD1, matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9) proteins. Results Compared with the NC group, NCI-H1299 cell viability, colony formation number, migration number, invasion number, the expression levels of CyclinD1, MMP2 and MMP9 proteins were decreased in the low, medium and high dose groups (P<0.05), and the expression of miR-100-5p was significantly increased (P<0.05). Compared with the miR-NC group, the expression of miR-100-5p was increased, the NCI-H1299 cell viability, colony formation number, migration number, invasion number, the expression of CyclinD1, MMP2 and MMP9 proteins decreased in the miR-100-5p group (P<0.05). Compared with the anti-miR-NC+ high-dose group, the expression of miR-100-5p was decreased, NCI-H1299 cell viability, colony formation number, migration number, invasion number, the expression of CyclinD1, MMP2 and MMP9 proteins increased in the anti-miR-100-5p+ high-dose group (P<0.05). Conclusion Lupeol inhibits the proliferation, migration and invasion of NSCLC cells NCI-H1299 by up-regulating miR-100-5p.

Key words: carcinoma, non-small-cell lung, lupeol, microRNAs, cell proliferation, cell movement, cyclin D1, matrix metalloproteinases, miR-100-5p

CLC Number:

R734.2

Figures/Tables 14

References 21

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组别	OD₄₅₀	细胞克隆数（个）	CyclinD1
NC组	1.01±0.10	113.00±8.26	0.85±0.09
低剂量组	0.90±0.09^a	84.00±7.15^a	0.73±0.07^a
中剂量组	0.73±0.07^ab	61.00±5.13^ab	0.53±0.05^ab
高剂量组	0.51±0.05^abc	50.00±4.02^abc	0.41±0.04^abc
F	67.047^＊＊	172.777^＊＊	81.965^＊＊

组别	OD₄₅₀	细胞克隆数（个）	CyclinD1
NC组	1.01±0.10	113.00±8.26	0.85±0.09
低剂量组	0.90±0.09^a	84.00±7.15^a	0.73±0.07^a
中剂量组	0.73±0.07^ab	61.00±5.13^ab	0.53±0.05^ab
高剂量组	0.51±0.05^abc	50.00±4.02^abc	0.41±0.04^abc
F	67.047^＊＊	172.777^＊＊	81.965^＊＊

组别	细胞迁移数（个）	细胞侵袭数（个）	MMP2	MMP9
NC组	154.00±12.25	136.00±13.32	0.83±0.08	0.75±0.06
低剂量组	127.00±10.13^a	101.00±10.03^a	0.70±0.07^a	0.62±0.06^a
中剂量组	100.00±9.24^ab	88.00±8.02^ab	0.57±0.05^ab	0.48±0.04^ab
高剂量组	62.00±5.16^abc	51.00±4.01^abc	0.42±0.04^abc	0.37±0.03^abc
F	152.245^＊＊	123.809^＊＊	72.156^＊＊	101.567^＊＊

组别	细胞迁移数（个）	细胞侵袭数（个）	MMP2	MMP9
NC组	154.00±12.25	136.00±13.32	0.83±0.08	0.75±0.06
低剂量组	127.00±10.13^a	101.00±10.03^a	0.70±0.07^a	0.62±0.06^a
中剂量组	100.00±9.24^ab	88.00±8.02^ab	0.57±0.05^ab	0.48±0.04^ab
高剂量组	62.00±5.16^abc	51.00±4.01^abc	0.42±0.04^abc	0.37±0.03^abc
F	152.245^＊＊	123.809^＊＊	72.156^＊＊	101.567^＊＊

组别	miR-100-5p	OD值	细胞克隆数（个）	细胞迁移数（个）	细胞侵袭数（个）	CyclinD1	MMP2	MMP9
miR-NC组	1.01±0.11	1.09±0.08	109.00±11.21	152.00±13.01	145.00±13.12	0.79±0.04	0.82±0.07	0.72±0.08
miR-100-5p组	2.68±0.34	0.69±0.08	56.00±4.12	85.00±6.33	68.00±5.11	0.52±0.06	0.56±0.07	0.45±0.06
t	14.020^＊＊	10.607^＊＊	13.313^＊＊	13.893^＊＊	16.406^＊＊	11.233^＊＊	7.879^＊＊	8.100^＊＊