Isovitexin regulates proliferation, migration and invasion of pancreatic cancer cells via the miR-339-5p/HSPA8 axis

doi:10.11958/20242056

Abstract

Abstract:

Objective To explore the biological behavior and mechanism of isovitexin (Isov) on pancreatic cancer cells. Methods Isov was used to treat the human normal pancreatic ductal epithelial cells HPDE and PC cell lines, and CCK-8 was used to detect the cell proliferation and calculate the half inhibitory concentration (IC₅₀). The PC cell line PANC-1 cells were grouped into the control group, the Isov group, the Isov+in-miR-NC group, the Isov+in-miR-339-5p group, the Isov+in-miR-339-5p+si-NC group and the Isov+in-miR-339-5p+si-HSPA8 group. The survival, migration and invasion of PANC-1 cells were detected by CCK-8, scratch healing assay and Transwell assay. Real time fluorescence quantitative PCR was used to detect the mRNA expression of miR-339-5p and heat shock protein family A member 8 (HSPA8) in PANC-1 cells. Western blot assay was used to detect protein HSPA8 expression in various groups of cells. Dual luciferase reporter gene was used to detect the targeting effect of miR-339-5p and HSPA8. A xenograft nude mouse model was used to determine the in vivo anticancer effects of Isov. Results Isov inhibited PC cell proliferation but had little cytotoxicity to HPDE cells. Isov could obviously reduce the survival rate and scratch healing rate of PANC-1 cells, reduce the number of invasive cells, up-regulate miR-339-5p expression and down-regulate HSPA8 mRNA and protein levels (P<0.05), while these effects were blocked by down-regulated miR-339-5p (P<0.05). In addition, HSPA8 was the target gene of miR-339-5p, and knockdown of HSPA8 reversed the regulatory effect of Isov on the malignant biological behavior of PANC-1 cells. In vivo studies confirmed that after Isov treatment, the tumor volume and weight of nude mice decreased, the expression of miR-339-5p was increased and the expression of HSPA8 mRNA was decreased (P<0.05). Conclusion Isov may inhibit the proliferation, migration and invasion of PC cells through the miR-339-5p/HSPA8 axis.

Key words: vitexin, pancreatic neoplasms, cell proliferation, HSPA8, miR-339-5p

CLC Number:

R735.9

Figures/Tables 9

References 23

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细胞类型	0 mmol/L	5 mmol/L	10 mmol/L	20 mmol/L	40 mmol/L	F
HPDE	100.00±0.00	108.36±9.79	104.37±8.76	98.65±7.65	92.32±8.15	1.853
AsPC-1	100.00±0.00	87.65±7.12	72.24±5.67^ABe	59.23±4.69^ABCe	43.12±4.62^ABCDe	60.181^＊＊
PANC-1	100.00±0.00	83.32±8.24^A	67.15±7.05^ABe	52.12±5.08^ABCe	35.18±4.13^ABCDe	60.480^＊＊
SW1990	100.00±0.00	89.17±8.25	75.24±6.80^Ae	62.13±5.62^ABe	47.12±4.23^ABCe	40.493^＊＊
F		5.237^＊＊	16.440^＊＊	37.790^＊＊	64.791^＊＊

细胞类型	0 mmol/L	5 mmol/L	10 mmol/L	20 mmol/L	40 mmol/L	F
HPDE	100.00±0.00	108.36±9.79	104.37±8.76	98.65±7.65	92.32±8.15	1.853
AsPC-1	100.00±0.00	87.65±7.12	72.24±5.67^ABe	59.23±4.69^ABCe	43.12±4.62^ABCDe	60.181^＊＊
PANC-1	100.00±0.00	83.32±8.24^A	67.15±7.05^ABe	52.12±5.08^ABCe	35.18±4.13^ABCDe	60.480^＊＊
SW1990	100.00±0.00	89.17±8.25	75.24±6.80^Ae	62.13±5.62^ABe	47.12±4.23^ABCe	40.493^＊＊
F		5.237^＊＊	16.440^＊＊	37.790^＊＊	64.791^＊＊

组别	细胞存活率/%	划痕愈合率/%	侵袭细胞数量/个	miR-339-5p	HSPA8 mRNA	HSPA8蛋白
Control组	100.00±0.00	68.54±6.25	159.65±12.36	1.02±0.05	1.03±0.04	0.85±0.05
Isov组	54.32±5.23^a	21.15±2.45^a	56.26±5.64^a	2.39±0.36^a	0.32±0.03^a	0.20±0.03^a
Isov+in-miR-NC组	51.36±4.62	23.36±3.05	61.23±6.28	2.05±0.30	0.30±0.02	0.25±0.04
Isov+in-miR-339-5p组	75.23±7.36^b	48.36±4.12^b	97.54±10.13^b	1.62±0.14^b	0.68±0.04^b	0.62±0.05^b
Isov+in-miR-339-5p+si-NC组	80.13±6.25	51.23±5.03	108.65±9.54	1.70±0.15	0.70±0.06	0.65±0.04
Isov+in-miR-339-5p+si-HSPA8组	63.12±5.45^c	37.54±3.25^c	81.36±7.65^c	1.73±0.16	0.50±0.08^c	0.40±0.05^c
F	34.980^＊＊	54.791^＊＊	54.402^＊＊	13.070^＊＊	94.071^＊＊	99.850^＊＊

组别	细胞存活率/%	划痕愈合率/%	侵袭细胞数量/个	miR-339-5p	HSPA8 mRNA	HSPA8蛋白
Control组	100.00±0.00	68.54±6.25	159.65±12.36	1.02±0.05	1.03±0.04	0.85±0.05
Isov组	54.32±5.23^a	21.15±2.45^a	56.26±5.64^a	2.39±0.36^a	0.32±0.03^a	0.20±0.03^a
Isov+in-miR-NC组	51.36±4.62	23.36±3.05	61.23±6.28	2.05±0.30	0.30±0.02	0.25±0.04
Isov+in-miR-339-5p组	75.23±7.36^b	48.36±4.12^b	97.54±10.13^b	1.62±0.14^b	0.68±0.04^b	0.62±0.05^b
Isov+in-miR-339-5p+si-NC组	80.13±6.25	51.23±5.03	108.65±9.54	1.70±0.15	0.70±0.06	0.65±0.04
Isov+in-miR-339-5p+si-HSPA8组	63.12±5.45^c	37.54±3.25^c	81.36±7.65^c	1.73±0.16	0.50±0.08^c	0.40±0.05^c
F	34.980^＊＊	54.791^＊＊	54.402^＊＊	13.070^＊＊	94.071^＊＊	99.850^＊＊

组别	HSPA8-WT	HSPA8-MUT
miR-NC组	1.00±0.06	1.02±0.05
miR-339-5p组	0.38±0.04	0.95±0.07
t	14.891^＊＊	1.409