miR-152降低低密度脂蛋白受体表达对子宫内膜癌细胞增殖和侵袭的抑制作用

doi:10.13481/j.1671-587X.20220306

Abstract

Abstract:

Objective： To investigate the expression and role of microRNA-152 （miR-152） in endometrial carcinoma （EC） tissue， and to provide the basis for study of pathogenesis and targeted treatment of EC.

Methods

The cancer tissues of 21 patients with EC （tumor group） and 39 patients with non-EC （control group） and human endometrial cancer RL95-2 cells and 293T cells were selected as the subjects. The clinical data such as age， height， body mass （BM）， body mass index （BMI）， systolic blood pressure （SBP）， diastolic blood pressure （DBP）， waist circumference （WC） and abdominal circumference （AC） of the patients in tumor group and control group were collected. The levels of triglyceride（TG），total cholesterol （TC）， low-density lipoprotein （LDL）， high density lipoprotein （HDL）， Ki-67 and low-density lipoprotein receptor （LDLR） of the patients in two groups were measured.Bioinformatics method was used to predict the expression of target gene LDLR related to the proliferation and invasion downstream of miR-152. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of miR-152 in endometrial tissue of the patients in tumor group and control group， Western blotting method and immunohistochemistry were used to detect the expression levels of LDLR protein in endometrial tissue of the patients in tumor group and control group， and Person correlation analysis was used to analyze the correlations between the expression of LDLR mRNA and the general data， biochemical indexes of the patients and miR-152 expression. The RL95-2 cells were divided into empty plasmid group （transfected with miR-NC empty plasmid）， miR-152 group （transfected with miR-152-mimics）， miR-152-mimics+pcDNA3.1-vector group （simultaneously transfected with miR-152-mimics and pcDNA3.1-vector）， and miR-152-mimics+pcDNA3.1-LDLR group （simultaneously transfected with miR-152-mimics+pcDNA3.1-LDLR）. The proliferation rates of the EC cells in various groups were detected by CCK-8 method， and the number of invasive EC cells in various groups was detected by Transwell assay. The luciferase activities of the 293T cells in various groups were detected by double luciferase reporter gene experiment.

Results

The expression level of miR-152 in endometrial tissue of the patients in tumor group was significantly lower than that in control group （P<0.05）， and the expression levels of LDLR mRNA and protein were higher than those in control group （P<0.05 or P<0.01）. The results of dual-luciferase reporter gene assay showed that LDLR was the target gene of miR-152.The Person correlation analysis results showed that the expression of LDLR mRNA in endometrial tissue of the patients in tumor group had a positive correlation with Ki-67， BMI， TC， TG， BM，and WC （r=0.4490， r=0.4377， r=0.4472， r=0.4706， r=0.5882， r=0.5130， P<0.05）， and it was negatively correlated with the expression of miR-152（r=－0.4378， P<0.05）. Compared with empty plasmid group， the proliferation rate of the RL95-2 cells in miR-152 group was decreased （P<0.05）， and the number of invasive cells was decreased （P<0.05）； compared with miR-152-mimics+pcDNA3.1-vector group， the proliferation rate of the cells in miR-152-mimics+pcDNA3.1-LDLR group was increased （P<0.01）， and the number of invasive cells was increased （P<0.01）.

Conclusion

MiR-152 inhibits the proliferation and invasion of the EC cells by decreasing the expression of LDLR.

Key words: MicroRNA-152, Endometrial carcinoma, Low-density lipoprotein receptor, Cell proliferation, Cell invasion

CLC Number:

R737.33

Guowu WANG,Yuan YAO,Yu ZHANG,Na XU,Fang LIU. Inhibitory effect of miR-152 on proliferation and invasion of endometrial carcinoma cells by reducing low-density lipoprotein receptor expression[J].Journal of Jilin University(Medicine Edition), 2022, 48(3): 591-599.

Figures/Tables 13

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References 26

1	CROSBIE E J， KITSON S J， MCALPINE J N， et al. Endometrial cancer［J］. Lancet， 2022， 399（10333）： 1412-1428.
2	周琦，吴小华，刘继红，等. 子宫内膜癌诊断与治疗指南（第四版）［J］. 中国实用妇科与产科杂志， 2018， 34（8）： 880-886.
3	WANG H G， YANG Q J， LI J P， et al. microRNA-15a-5p inhibits endometrial carcinoma proliferation， invasion and migration via downregulation of VEGFA and inhibition of the Wnt/β-catenin signaling pathway［J］. Oncol Lett， 2021， 21（4）： 310.
4	GENG A Z， LUO L， REN F Y， et al. miR-29a-3p inhibits endometrial cancer cell proliferation， migration and invasion by targeting VEGFA/CD C42/PAK1［J］. BMC Cancer， 2021， 21（1）： 843.
5	SUN X M， HOU L T， QIU C P， et al. miR-501 promotes tumor proliferation and metastasis by targeting HOXD10 in endometrial cancer［J］. Cell Mol Biol Lett， 2021， 26（1）： 20.
6	CORREIA DE SOUSA M， GJORGJIEVA M， DOLICKA D， et al. Deciphering miRNAs’ action through miRNA editing［J］.Int J Mol Sci，2019，20（24）： E6249.
7	GE S K， WANG D， KONG Q L， et al. Function of miR-152 as a tumor suppressor in human breast cancer by targeting PIK3CA［J］. Oncol Res， 2017， 25（8）： 1363-1371.
8	LIU P H， WU L X， CHAND H， et al. Silencing of miR-152 contributes to DNMT1-mediated CpG methylation of the PTEN promoter in bladder cancer［J］. Life Sci， 2020， 261： 118311.
9	LI B， XIE Z S， LI B. miR-152 functions as a tumor suppressor in colorectal cancer by targeting PIK3R3［J］. Tumour Biol， 2016， 37（8）： 10075-10084.
10	黄雨欣，雷蕾，华克勤，辛卫娟.长链非编码RNA R P11-395G23.3在子宫内膜癌中的表达及其临床意义［J］.同济大学学报（医学版），2021，42（3）：321-326.
11	ZHAO YOU-BO， YUAN D Z， PAN J L， et al. Effect of progesterone-induced microRNA-152 on the proliferation of endometrial epithelial cells［J］. J Sichuan Univ Med Sci Ed， 2016， 47（2）： 147-151.
12	BJORNSSON E， GUNNARSDOTTIR K， HALLDORSSON G H， et al. Lifelong reduction in LDL （low-density lipoprotein） cholesterol due to a gain-of-function mutation in LDLR［J］. Circ Genom Precis Med， 2021， 14（1）： e003029.
13	FELDT M， MENARD J， ROSENDAHL A H， et al. The effect of statin treatment on intratumoral cholesterol levels and LDL receptor expression：a window-of-opportunity breast cancer trial［J］. Cancer Metab， 2020， 8（1）： 25.
14	NIU X L， NONG S R， GONG J Y， et al. miR-194 promotes hepatocellular carcinoma through negative regulation of CADM1［J］. Int J Clin Exp Pathol， 2020， 13（7）： 1518-1528.
15	LI X N， YANG H， YANG T. miR-122 inhibits hepatocarcinoma cell progression by targeting LMNB₂ ［J］. Oncol Res， 2020， 28（1）： 41-49.
16	SARVIZADEH M， MALEKSHAHI Z V， RAZI E， et al. microRNA： a new player in response to therapy for colorectal cancer［J］. J Cell Physiol， 2019， 234（6）： 8533-8540.
17	HUANG S L， TAN X H， HUANG Z C， et al. microRNA biomarkers in colorectal cancer liver metastasis［J］. J Cancer， 2018， 9（21）： 3867-3873.
18	LI L W， XIAO H Q， MA R， et al. miR-152 is involved in the proliferation and metastasis of ovarian cancer through repression of ERBB₃ ［J］. Int J Mol Med， 2018， 41（3）： 1529-1535.
19	XIANG Y， MA N， WANG D， et al. miR-152 and miR-185 co-contribute to ovarian cancer cells cisplatin sensitivity by targeting DNMT1 directly： a novel epigenetic therapy independent of decitabine［J］. Oncogene， 2014， 33（3）： 378-386.
20	ZHANG H Y， LU Y X， WANG S R， et al. microRNA-152 acts as a tumor suppressor microRNA by inhibiting krüppel-like factor 5 in human cervical cancer［J］. Oncol Res， 2019， 27（3）： 335-340.
21	HUANG S D， LI X H， ZHU H T. microRNA-152 targets phosphatase and tensin homolog to inhibit apoptosis and promote cell migration of nasopharyngeal carcinoma cells［J］. Med Sci Monit，2016，22：4330-4337.
22	JEON H， BLACKLOW S C. Structure and physiologic function of the low-density lipoprotein receptor［J］. Annu Rev Biochem， 2005， 74： 535-562.
23	WILLNOW T E， NYKJAER A， HERZ J. Lipoprotein receptors： new roles for ancient proteins［J］. Nat Cell Biol， 1999， 1（6）： E157-E162.
24	STOPSACK K H， GERKE T A， ANDRÉN O， et al. Cholesterol uptake and regulation in high-grade and lethal prostate cancers［J］.Carcinogenesis，2017，38（8）：806-811.
25	SUN Y， FENG Y， ZHANG G Q， et al. The endonuclease APE1 processes miR-92b formation， thereby regulating expression of the tumor suppressor LDLR in cervical cancer cells［J］. Ther Adv Med Oncol， 2019， 11： 1758835919855859.
26	CHEN Z Y， CHEN L， SUN B， et al. LDLR inhibition promotes hepatocellular carcinoma proliferation and metastasis by elevating intracellular cholesterol synthesis through the MEK/ERK signaling pathway［J］. Mol Metab， 2021， 51： 101230.

Group	n	Age (years)	BMI (kg·m^－2)	BM (m/kg)	WC (l/cm)	SBP (P/mmHg)
Normal	39	59.21±5.41	22.68±2.41	59.10±6.81	90.15±8.18	123.70±14.15
Tumor	21	60.62±7.32	26.73±3.51	64.48±8.44	97.00±9.51	125.30±16.78
P		0.785 0	0.000 2	0.048 6	0.003 5	0.865 9
Group	n	TC [c_B/(mmol·L^－1]	TG [c_B/(mmol·L^－1]	HDL [c_B/(mmol·L^－1]	LDL [c_B/(mmol·L^－1]	DBP (P/mmHg)
Normal	39	3.65±0.75	1.35±0.51	1.23±0.80	2.13±0.73	70.59±9.58
Tumor	21	4.17±0.94	1.54±0.40	1.11±0.59	3.15±0.93	70.52±8.48
P		0.046 4	0.004 4	0.329 3	0.002 2	0.815 1

Target detail	Target rank	Target score	miRNA name	Gene symbol	Gene description
Details	1	100	has-miR-152-3P	SOS2	SOS Ras/Rho guanine nucleotide exchange factor 2
Details	2	100	has-miR-152-3P	CDK19	Cyclin dependent kinase 19
Details	3	99	has-miR-152-3P	UBE2D3	Ubiquitin conjugating enzyme E2 D3
Details	4	99	has-miR-152-3P	RPS6KA5	Ribosomal protein S6 kinase A5
Details	5	99	has-miR-152-3P	HOMER1	Homer scaffold protein 1
Details	6	99	has-miR-152-3P	BCL2L11	BCL2 like 11
Details	7	99	has-miR-152-3P	LDLR	Low density lipoprotein receptor
Details	8	99	has-miR-152-3P	CBLB	Cbl proto-oncogene B
Details	9	99	has-miR-152-3P	MEOX2	Mesenchyme homeobox 2
Details	10	99	has-miR-152-3P	KMT2A	Lysine wethyltransferase 2A
Details	11	99	has-miR-152-3P	WDR47	WD repeat domain 47
Details	12	99	has-miR-152-3P	NPTN	Neuroplastin

Correlation	LDLR mRNA
Correlation	r	P
Ki-67	0.449 0	0.041 2
BMI	0.437 7	0.047 2
TC	0.447 2	0.042 1
TG	0.470 6	0.031 3
BM	0.513 0	0.017 4
WC	0.588 2	0.005 0

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Inhibitory effect of miR-152 on proliferation and invasion of endometrial carcinoma cells by reducing low-density lipoprotein receptor expression

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