石榴皮多酚调控miR-138-5p/HIF-1α通路对结肠癌细胞恶性生物学行为的影响

doi:10.3969/j.issn.1006-5725.2025.05.009

Abstract

Abstract:

Objective To investigate the impact of pomegranate peel polyphenols （PPP） on the malignant biological behavior of colon cancer cells through modulation of the miR?138?5p/hypoxia?inducible factor?1α （HIF?1α） pathway. Methods Quantitative real?time PCR （qRT?PCR） was employed to measure the expression levels of miR?138?5p and HIF?1α mRNA in the normal colon epithelial cell line FHC and three colorectal cancer cell lines： SW480， HCT116， and Caco?2. SW480 cells were divided into six groups： a blank control group， a negative control （mimics NC） group， a miR?138?5p mimics group， three different concentrations of PPP treatment groups （0.5 mg/mL， 1 mg/mL， and 2 mg/mL）， a PPP + inhibitor NC group at 2 mg/mL， and a PPP + miR?138?5p inhibitor group at 2 mg/mL. The effects on cell proliferation， invasion， and migration， as well as changes in apoptosis and related proteins including B?cell lymphoma 2 （Bcl?2）， migration invasion enhancer 1 （MIEN1）， and Cyclin D1， were evaluated separately. Additionally， the targeting relationship between miR?138?5p and HIF?1α was validated. The expression levels of miR?138?5p， HIF?1α mRNA， and protein were assessed in each experimental group. Results The expression levels of miR?138?5p were highest in FHC cells and lowest in SW 480 cells， while the expression levels of HIF?1α mRNA showed an opposite trend， being lowest in FHC cells and highest in SW 480 cells （P < 0.05）. Compared with the control group， different concentrations of PPP significantly promoted cell apoptosis， upregulated miR?138?5p expression， inhibited cell proliferation， invasion， and migration， and downregulated the expression of HIF?1α mRNA， Bcl?2， MIEN1， CyclinD1， and HIF?1α protein， with significant differences between groups （P < 0.05）. Compared with the mimics NC group， the miR?138?5p mimics group significantly enhanced cell apoptosis， upregulated miR?138?5p expression， inhibited cell proliferation， invasion， and migration， and downregulated the expression of HIF?1α mRNA， Bcl?2， MIEN1， CyclinD1， and HIF?1α protein （P < 0.05）. Compared with the 2 mg/mL PPP + inhibitor NC group， the 2 mg/mL PPP + miR?138?5p inhibitor group significantly suppressed cell apoptosis， downregulated miR?138?5p expression， promoted cell proliferation， invasion， and migration， and upregulated the expression of HIF?1α mRNA， Bcl?2， MIEN1， CyclinD1， and HIF?1α protein （P < 0.05）. These results indicate a targeted relationship between miR?138?5p and HIF?1α （P < 0.05）. Conclusion PPP inhibits the malignant biological behavior of colon cancer cells through upregulation of the miR?138?5p/HIF?1α pathway.

Key words: pomegranate peel polyphenols, miR-138-5p/HIF-1α pathway, colon cancer, proliferation, apoptosis, invasion, migration

CLC Number:

R285.5

Hongyan BIAN,Shu ZHANG,Shanshan MENG,Ying WEI. Effect of pomegranate peel polyphenols on the malignant biological behavior of colon cancer cells by regulating the miR⁃138⁃5p/HIF⁃1α pathway[J]. The Journal of Practical Medicine, 2025, 41(5): 676-682.

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引物名称	方向	序列（5′-3′）
miR-138-5p	F	UUAAUGCUAAUCGUGAUAGGGGU
miR-138-5p	R	ACCCCUAUCACGAUUAGCAUUAA
HIF-1α	F	TACAGTACTGTGATAACTGAA
HIF-1α	R	GCAGGGTCCGAGGTATTC
β-actin	F	CGAGCCACATCGCTCAGACA
β-actin	R	GTGGTGAAGACGCCAGTGGA
U6	F	GAGAAGATTAGCATGGCCCCT
U6	R	ATTTCTCGATTTGTGCGTGTCA

组别	miR-138-5p	HIF-1α mRNA
FHC细胞	1.06 ± 0.11	0.94 ± 0.10
Caco-2细胞	0.63 ± 0.07^?	1.56 ± 0.17^?
HCT116细胞	0.66 ± 0.07^?	1.61 ± 0.18^?
SW480细胞	0.42 ± 0.05^?	1.88 ± 0.20^?
F值	70.254	34.046
P值	< 0.001	< 0.001

组别	增殖率/%
空白组	94.05 ± 3.52
mimics NC组	95.11 ± 3.58
miR-138-5p mimics组	71.05 ± 3.15^*#
0.5 mg/mL PPP组	72.04 ± 3.18^*
1 mg/mL PPP组	50.21 ± 3.09^*△
2 mg/mL PPP组	35.51 ± 3.02^*△▲
2 mg/mL PPP + inhibitor NC组	36.02 ± 3.05
2 mg/mL PPP + miR-138-5p inhibitor组	70.61 ± 3.13^▽
F值	313.389
P值	< 0.001

组别	侵袭数	迁移数
空白组	152.31 ± 16.11	146.25 ± 15.24
mimics NC组	149.85 ± 15.02	145.84 ± 14.75
miR-138-5p mimics组	110.24 ± 11.42^*#	99.85 ± 10.01^*#
0.5 mg/mL PPP组	114.03 ± 12.06^*	100.52 ± 10.52^*
1 mg/mL PPP组	82.51 ± 9.02^*△	78.53 ± 7.94^*△
2 mg/mL PPP组	57.64 ± 5.83^*△▲	42.06 ± 4.33^*△▲
2 mg/mL PPP + inhibitor NC组	56.21 ± 5.79	41.86 ± 4.25
2 mg/mL PPP + miR-138-5p inhibitor组	109.85 ± 11.21^▽	98.56 ± 10.03^▽
F值	62.939	88.947
P值	< 0.001	< 0.001

组别	凋亡率
空白组	7.02 ± 0.72
mimics NC组	6.86 ± 0.71
miR-138-5p mimics组	12.75 ± 1.31^*#
0.5 mg/mL PPP组	12.52 ± 1.31^*
1 mg/mL PPP组	22.05 ± 2.24^*△
2 mg/mL PPP组	35.06 ± 3.58^*△▲
2 mg/mL PPP+inhibitor NC组	35.42 ± 3.62
2 mg/mL PPP+miR-138-5p inhibitor组	13.06 ± 1.33^▽
F值	172.986
P值	< 0.001

Effect of pomegranate peel polyphenols on the malignant biological behavior of colon cancer cells by regulating the miR⁃138⁃5p/HIF⁃1α pathway

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组别	miR-138-5p	HIF-1α mRNA
空白组	1.01 ± 0.11	0.98 ± 0.10
mimics NC组	0.98 ± 0.10	1.05 ± 0.11
miR-138-5p mimics组	1.51 ± 0.16^*#	0.59 ± 0.07^*#
0.5 mg/mL PPP组	1.45 ± 0.16^*	0.65 ± 0.07^*
1 mg/mL PPP组	1.86 ± 0.19^*△	0.42 ± 0.05^*△
2 mg/mL PPP组	2.54 ± 0.26^*△▲	0.23 ± 0.03^*△▲
2 mg/mL PPP + inhibitor NC组	2.61 ± 0.27	0.28 ± 0.03
2 mg/mL PPP + miR-138-5p inhibitor组	1.85 ± 0.20^▽	0.55 ± 0.06^▽
F值	63.140	107.403
P值	< 0.001	< 0.001

组别	荧光素酶相对活性
mimics NC+HIF-1α WT组	1.01 ± 0.11
miR-138-5p mimics+HIF-1α WT组	0.55 ± 0.06^*
mimics NC+HIF-1α MUT组	1.05 ± 0.11
miR-138-5p mimics+HIF-1α MUT组	1.07 ± 0.11
F值	36.972
P值	<0.001

组别	Bcl-2/β-actin	MIEN1/β-actin	CyclinD1/β-actin	HIF-1α/β-actin
空白组	1.27 ± 0.14	1.02 ± 0.12	1.48 ± 0.16	1.31 ± 0.14
mimics NC组	1.31 ± 0.14	1.01 ± 0.11	1.39 ± 0.15	1.29 ± 0.13
miR-138-5p mimics组	0.80 ± 0.09^*#	0.69 ± 0.07^*#	0.98 ± 0.10^*#	0.79 ± 0.08^*#
0.5 mg/mL PPP组	0.83 ± 0.09^*	0.72 ± 0.08^*	1.01 ± 0.11^*	0.89 ± 0.09^*
1 mg/mL PPP组	0.52 ± 0.06^*△	0.47 ± 0.05^*△	0.76 ± 0.08^*△	0.63 ± 0.07^*△
2 mg/mL PPP组	0.24 ± 0.03^*△▲	0.22 ± 0.03^*△▲	0.43 ± 0.05^*△▲	0.38 ± 0.04^*△▲
2 mg/mL PPP+inhibitor NC组	0.22 ± 0.03	0.28 ± 0.03	0.41 ± 0.05	0.34 ± 0.04
2 mg/mL PPP+miR-138-5p inhibitor组	0.75 ± 0.08^▽	0.65 ± 0.07^▽	0.93 ± 0.10^▽	0.82 ± 0.09^▽
F值	121.056	91.149	80.248	94.427
P值	< 0.001	< 0.001	< 0.001	< 0.001

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