Expression And Clinical Significance Of LncRNA ANRIL In Human Endometrial Carcinoma

Objective: To detect the expression level of long non-coding RNA(lncRNA)ANRIL(antisense long non-coding RNA in the INK4 locus)molecule in endometrial carcinoma tissues,normal tissues and human endometrial carcinoma HEC-1A cells,and si RNA-ANRIL(small interfering RNA of ANRIL)was employed to negatively regulate the gene expression of ANRIL,then observed the influence on the proliferation of human endometrial carcinoma cell line HEC-1A and further analyzed its expression level and the relationship between the clinical pathology characteristic parameters and its clinical significance.Methods:1 Eighty cases of fresh human endometrium were collected from 2009 April to 2015 November at the First Hospital of Hebei Medical University including 40 cases normal endometrium tissues as control group,and 40 cases endometrial carcinoma tissues(endometrioid adeno carcinoma 32 cases,adenocarcinoma with squamous differentiation 3 cases,serous adenocarcinoma 3 cases,mucinous carcinoma 1 case and clear cell carcinoma 1 case),as experimental group.2 Total RNA were obtained by Trizol.and Revert Aid First strand c DNA Synthesis Kit was used to obtain c DNA.3 Real-time PCR was employed to detect the expression of ANRIL in tissues and cells.4 Human endometrial carcinoma HEC-1A(moderately differentiated)cells were cultured,subcultured and cryopreserved for follow-up cytological experiments.5 In this research,the cells were transfected by liposome(Lipofectamine 2000 transfection)mediated which allowed foreign genes si RNAs targeting ANRIL(si RNA-ANRIL)or negative controls(si RNA-NC)enter cells by membrane fusion and these foreign genes may triggered their complementary target messenger RNA(m RNA)specific degradation then silenced target gene ANRIL expression.6 Clony formation assay and crystal violet assay were used to detect the effect of si RNA-ANRIL(experimental group)on cell proliferation in human endometrial carcinoma cells,compared with si RNA-NC(negative control group)and untreated group(blank control group).Results:1 The relative expression levels of long non-coding RNA ANRIL in endometrial carcinoma tissues was significant higher than in normal endometrium tissues(P<0.05),and the expression of ANRIL was also higher in histologic grade ? samples than histologic grade ?,? samples.However,the results revealed that the expression of ANRIL was not related to ages,FIGO stages,pelvic lymph node metastasis,estrogen receptor expression,progestin receptor expression and with/without diabetes or hypertension.2 In vitro,si RNA-ANRIL were transfected into HEC-1A cells,as the experimental group,the expression level as mean±SD were significantly reduced compared with negative control group and blank control group(0.323738±0.037062,0.4391±0.06164,0.406675±0.055332,P<0.05).Conclusion:1 The significantly higher expression of lncRNA ANRIL in carcinoma tissues than in normal tissues.2 The higher expression of ANRIL in histologic grade ? tissues than histologic grade ?,? tissues,which indicated ANRIL may play the part of cancer-promoting genetic in the development and progression of endometrial adenocarcinoma.3 LncRNA ANRIL was down-regulated by si RNA-ANRIL in human endometrial carcinoma HEC-1A cells,si RNA-ANRIL can obviously inhibit cell proliferation,which illustrated that ANRIL probably targeted modulate human endometrial carcinoma cell proliferation.In summary,according to the result of the experiment,we may carefully conjecture that some change of epigenetic modifications may promote the expression of ANRIL,and the overexpression of ANRIL may play a cancer-promoting gene role in the development of endometrial carcinoma.