微小RNA-365和靶向E74样受体4在宫颈癌细胞的增殖及其临床意义

doi:10.3881/j.issn.1000-503X.10613

Acta Academiae Medicinae Sinica ›› 2019, Vol. 41 ›› Issue (2): 220-227.doi: 10.3881/j.issn.1000-503X.10613

• Original Articles • Previous Articles Next Articles

Proliferation of MicroRNA-365 and E74-like Factor 4 in Cervical Cancer Cells and Its Clinical Significance

GUO Ying¹,MA Dong¹,JIA Shifeng²,LIU Jia¹,FAN Shaobei¹,ZHANG Meng¹,SHI Linran¹,JIANG Lili¹,SHI Juexin¹,WANG Haiqiu³,ZHENG Huanyu³,LI Ou³()

1 Graduate School,North China University of Science and Technology,Tangshan,Hebei 063200,China
2 Department of Surgical Oncology,Affiliated Hospital of North China University of Science and Technology,Tangshan,Hebei 063000,China
3 Department of Obstetrics and Gynecology,Workers’ Hospital of Tangshan,Tangshan,Hebei 063000,China;

Received:2018-06-08 Online:2019-04-28 Published:2019-05-07
Contact: Ou LI E-mail:tsliou@163.com
Supported by:
Supported by the National Natural Sciences Foundation of China(81541149);the Youth National Natural Sciences Foundation of Hebei(H 2016209046);the Key Medical Sciences Research of Hebei(20180729)

Abstract

Abstract:

Objective To investigate the expressions,roles,and clinical significance of microRNA-365(miR-365)and E74-like factor 4(ELF4)in cervical cancer.Methods The expressions of miR-365 in normal cervical tissues(n=34),cervical intraepithelial neoplasia Ⅰ(CIN Ⅰ)(n=31),cervical intraepithelial neoplasiaⅡ-Ⅲ(CIN Ⅱ-Ⅲ)(n=37),squamous cell carcinoma of the cervix(SCC)(n=33),and three cervical cancer cell lines(C33A cells,Hela cells,and SiHa cells)were detected by real-time quantitative polymerase chain reaction(qPCR).Bioinformatic prediction and luciferase reporter gene assay were performed to verify whether ELF4 was a direct target of miR-365.Western blot and immunohistochemistry were used to detect ELF4 expression in cervical cancer cells and in different pathological cervix tissues.CCK8 assay was used to detect the effect of overexpression or inhibition of miR-365 on the proliferation of cervical cancer cells at different time points.The relationships among the miR-365 expression,ELF4 expression,and clinicopathological parameters of cervical cancer were analyzed by correlation analysis.Results qPCR results showed that compared with the normal cervical cell HcerEpic,the expressions of miR-365 in CINⅠ,CINⅡ-Ⅲ,and cervical cancer tissues gradually decreased with the increased pathologic grade,and its expressions also decreased in different cervical cancer cell lines.The luciferase reporter gene assay confirmed that ELF4 was the direct target of miR-365.Western blot showed that the expression of ELF4 increased in all three cervical cancer cell lines compared with normal cervical epidermal cell(P=0.013,P=0.002,P=0.004).Immunohistochemistry showed that ELF4 expression was up-regulated in CIN and cervical cancer tissues.CCK8 assay showed that overexpression of miR-365 inhibited cell proliferation,while inhibition of miR-365 promoted the proliferation of three cervical cancer cells(P<0.05).Further analysis confirmed that there was a negative correlation between the expression levels of miR-365 and ELF4 in CINⅡ-Ⅲ and SCC(r=-0.351,P=0.045;r=-0.349,P=0.035).Clinical analysis showed that the expressions of both miR-365 and ELF4 were correlated with tumor size,pathological grade,and clinical stage in SCC(all P < 0.05).Conclusion The decreased expression of miR-365 in human cervical cancer cells relieves its inhibitory effect on ELF4,which promotes the proliferation of cervical cancer cells and the formation of tumor.

Key words: cervical cancer, microRNA-365, E74-like factor 4

CLC Number:

R737.33

GUO Ying,MA Dong,JIA Shifeng,LIU Jia,FAN Shaobei,ZHANG Meng,SHI Linran,JIANG Lili,SHI Juexin,WANG Haiqiu,ZHENG Huanyu,LI Ou. Proliferation of MicroRNA-365 and E74-like Factor 4 in Cervical Cancer Cells and Its Clinical Significance[J].Acta Academiae Medicinae Sinica, 2019, 41(2): 220-227.

Figures/Tables 8

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Table 1

Correlation between miR-365 expression and clinicopathological features of squamous cell carcinoma of the cervix tissues"

临床病理特征 Clinicopathological features	低表达Low expression	高表达High expression	t/χ²	P
年龄( $x ?$ ±s,岁)Age( $x ?$ ±s,years)	53.17±4.11	50.47±4.52	0.929	0.363
肿瘤大小Tumor size[n(%)]			4.891	0.027
≤4cm	40(30.8)	9(69.2)
>4cm	14(70.0)	6(30.0)
病理分级Pathological grade[n(%)]			6.906	0.009
高、中分化Well and moderately-differentiated	16(69.6)	7(30.4)
低分化Poorly-differentiated	2(20.0)	8(80.0)
临床分期Clinical stage[n(%)]			6.639	0.010
Ⅰ期ⅠStage	15(71.4)	6(28.6)
Ⅱ期ⅡStage	3(25.0)	9(75.0)

Table 1

Table 2

Correlation between ELF4 mRNA expression and clinicopathological features of squamous cell carcinoma of the cervix tissue"

临床病理特征 Clinicopathological features	低表达Low expression	高表达High expression	t/χ²	P
年龄( $x ?$ ±s,岁)Age( $x ?$ ±s,years)	53.85±4.39	50.70±4.12	1.655	0.120
肿瘤大小Tumor size[n(%)]			7.998	0.005
≤4cm	9(69.3)	4(30.8)
>4cm	4(20.0)	16(80.0)
病理分级Pathological grade[n(%)]			5.192	0.023
高、中分化Well and moderately-differentiated	12(52.2)	11(47.8)
低分化Poorly-differentiated	1(10.0)	9(90.0)
临床分期Clinical stage[n(%)]			4.080	0.043
Ⅰ期ⅠStage	11(52.4)	10(47.6)
Ⅱ期ⅡStage	2(16.7)	10(83.3)

Table 2

References 20

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Proliferation of MicroRNA-365 and E74-like Factor 4 in Cervical Cancer Cells and Its Clinical Significance

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