Impacts of Casticin on biological behavior of gastric cancer cells by regulating miR-378/PRRX1 axis

doi:10.11958/20222053

Abstract

Abstract:

Objective To explore effects of Casticin (Cas) on the proliferation, invasion, migration and microRNA-378 (miR-378)/paired related homeobox 1 (PRRX1) axis of gastric cancer (GC) SGC-7901 cells. Methods SGC-7901 cells were treated with Cas at different concentrations (0, 5, 10, 20, 40 and 80 μmol/L) for 48 h, and the cell viability of SGC-7901 was detected by tetrazole salt (MTT) method. SGC-7901 cells at logarithmic growth stage were divided into 7 groups: the control group, the Cas low concentration group (10 μmol/L), the Cas medium concentration group (20 μmol/L), the Cas high concentration group (40 μmol/L), the Cas high concentration+miR-378 small interfering RNA (siRNA) group (40 μmol/L Cas+miR-378 siRNA), the Cas high concentration+PRRX1 group (40 μmol/L Cas+PRRX1 overexpressed plasmid) and the Cas high concentration+miR-378 siRNA+PRRX1 group (40 μmol/L Cas+miR-378 siRNA+PRRX1 overexpression plasmid). Cell activity, colony formation number, apoptosis rate, invasion and migration ability of SGC-7901 cells were determined by MTT assay, colony formation assay, flow cytometry, Transwell chamber assay and scratch assay, respectively. Expression levels of miR-378 and PRRX1 protein in SGC-7901 cells were detected by fluorescence quantitative PCR and Western blot assay. Double luciferase assay verified the targeting relationship between miR-378 and PRRX1. Xenografted SGC-7901 cells were xenografted into nude mice to verify the anticancer activity of Cas and its effect on the expression of miR-378. Results The cell viability of SGC-7901 cells in Cas 10, 20, 40 and 80 μmol/L groups decreased in a concentration-dependent manner (P<0.05), and 10, 20 and 40 μmol/L Cas were selected as the follow-up study concentration. Compared with the control group, the cell viability, colony formation number, invasion cell number, scratch healing rate and PRRX1 protein expression level of SGC-7901 cells in the Cas low, medium and high concentration groups were decreased successively (P<0.05), while the apoptosis rate and the expression level of miR-378 were increased successively (P<0.05). Compared with Cas high concentration group, the cell viability, colony formation number, invasion cell number, scratch healing rate and PRRX1 protein expression levels were increased in the Cas high concentration+miR-378 siRNA group and the Cas high concentration+PRRX1 group, and the apoptosis rate was decreased (P<0.05). The expression level of miR-378 in SGC-7901 cells was decreased in the Cas high concentration+miR-378 siRNA group (P<0.05). Compared with the Cas high concentration+miR-378 siRNA group, the expression level of miR-378 in SGC-7901 cells was increased in the Cas high concentration+PRRX1 group (P<0.05). Compared with the Cas high concentration+miR-378 siRNA group and the Cas high concentration+PRRX1 group, the cell viability, colony formation number, invasion cell number, scratch healing rate and PRRX1 protein expression levels were increased in the Cas high concentration+miR-378 siRNA+PRRX1 group (P<0.05), and the apoptosis rate was decreased (P<0.05). In SGC-7901 cells, miR-378 could target the expression of PRRX1. Cas inhibited the growth of transplanted tumor and promoted the expression of miR-378 in nude mice (P<0.05). Conclusion Cas may inhibit the proliferation, invasion and migration of SGC-7901 cells by regulating the miR-378/PRRX1 axis.

Key words: viticis fructus, stomach neoplasms, microRNAs, cell proliferation, cell movement, tumor infiltration, homeodomain proteins

CLC Number:

R285.5，R735.2

Figures/Tables 12

References 21

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组别	细胞活力/%	集落形成数量/个	细胞凋亡率/%
对照组	100.00±0.00	124.56±10.25	4.01±0.24
Cas低浓度组	81.12±7.62^a	88.26±7.06^a	8.20±0.85^a
Cas中浓度组	68.24±6.53^ab	74.15±6.85^ab	15.83±1.77^ab
Cas高浓度组	52.03±5.07^abc	60.24±5.12^abc	26.57±2.19^abc
Cas高浓度+miR- 378 siRNA组	77.04±6.80^d	86.35±7.68^d	8.64±0.93^d
Cas高浓度+ PRRX1组	80.28±7.71^d	90.36±9.34^d	9.02±1.05^d
Cas高浓度+miR- 378 siRNA+ PRRX1组	93.40±8.55^ef	115.28±10.17^ef	5.10±0.47^ef
F	34.436^＊＊	43.591^＊＊	235.574^＊＊

组别	细胞活力/%	集落形成数量/个	细胞凋亡率/%
对照组	100.00±0.00	124.56±10.25	4.01±0.24
Cas低浓度组	81.12±7.62^a	88.26±7.06^a	8.20±0.85^a
Cas中浓度组	68.24±6.53^ab	74.15±6.85^ab	15.83±1.77^ab
Cas高浓度组	52.03±5.07^abc	60.24±5.12^abc	26.57±2.19^abc
Cas高浓度+miR- 378 siRNA组	77.04±6.80^d	86.35±7.68^d	8.64±0.93^d
Cas高浓度+ PRRX1组	80.28±7.71^d	90.36±9.34^d	9.02±1.05^d
Cas高浓度+miR- 378 siRNA+ PRRX1组	93.40±8.55^ef	115.28±10.17^ef	5.10±0.47^ef
F	34.436^＊＊	43.591^＊＊	235.574^＊＊

组别	侵袭细胞数量/ （个/视野）	划痕愈合率/%
对照组	240.15±22.15	65.34±5.89
Cas低浓度组	191.34±18.79^a	42.29±4.05^a
Cas中浓度组	151.62±13.49^ab	34.80±3.15^ab
Cas高浓度组	114.39±10.36^abc	24.33±2.08^abc
Cas高浓度+miR-378 siRNA组	163.69±15.21^d	40.12±4.08^d
Cas高浓度+PRRX1组	169.62±17.05^d	41.61±5.07^d
Cas高浓度+miR-378 siRNA+ PRRX1组	228.75±20.34^ef	57.35±5.64^ef
F	39.274^＊＊	56.610^＊＊

组别	侵袭细胞数量/ （个/视野）	划痕愈合率/%
对照组	240.15±22.15	65.34±5.89
Cas低浓度组	191.34±18.79^a	42.29±4.05^a
Cas中浓度组	151.62±13.49^ab	34.80±3.15^ab
Cas高浓度组	114.39±10.36^abc	24.33±2.08^abc
Cas高浓度+miR-378 siRNA组	163.69±15.21^d	40.12±4.08^d
Cas高浓度+PRRX1组	169.62±17.05^d	41.61±5.07^d
Cas高浓度+miR-378 siRNA+ PRRX1组	228.75±20.34^ef	57.35±5.64^ef
F	39.274^＊＊	56.610^＊＊

组别	miR-378	PRRX1蛋白（/GAPDH）
对照组	1.02±0.05	0.85±0.07
Cas低浓度组	1.58±0.10^a	0.68±0.06^a
Cas中浓度组	1.96±0.18^ab	0.57±0.06^ab
Cas高浓度组	2.34±0.21^abc	0.46±0.04^abc
Cas高浓度+miR-378 siRNA组	1.56±0.14^d	0.58±0.06^d
Cas高浓度+PRRX1组	2.31±0.20^e	0.62±0.07^d
Cas高浓度+miR-378 siRNA+ PRRX1组	1.54±0.16^f	0.79±0.06^ef
F	54.139^＊＊	29.519^＊＊