| Background and Aims: Currently, researches about the natural biological agents to treat tumor have become one of the most promising ways to the field of cancer treatment. Many researches have been done in that how the resveratrol inbit in cancer did, but it is uncertain in Endometrial Carcinoma. This study aimed to investigate the effections of microRNA-181a(miR-181a) and resveratrol in A Nude Mouse Model of Endometrial Carcinoma cells (ishikawa) Aim to verify that the effects in vivo are consistent with in vitro, and provide new ideas for clinical treatment.Methods:1.20 primary EC tumor tissue specimens were obtained from EC patients and 13 normal endometrial tissue samples were collected from women who underwent hysterectomy in Nanjing Drum Tower Hospital. miR-181a and SIRT 1 expression in different tissues were tested by Realtime-PCR and Western blot analysis.2. Following the construction of nude mouse tumor xenograft model with Ishikawa cells, all of the injected nude mice with tumor burden were randomly divided into six groups with 5 mice in each group when the volume of xenografts reached 50mm3(approximatery 2 weeks):A group (1xPBS intratumoral injection,10% CMC in gavage); B group (1xPBS intratumoral injection, resvertrol in gavage); C group (Ad-miR-181a intratumoral injection,10% CMC in gavage); D group (Ad-miR-181a intratumoral injection, resvertrol in gavage); E group(laz-miR-181a intratumoral injection,10% CMC in gavage); F group (laz-miR-181a intratumoral injection, resvertrol in gavage); The specific dose:injection by 1xPBS (10ul-mouse-1), Ad-miR-181a (2*10^8-mouse-1 10ul-mouse-1), laz-miR-181a (2*10^8-mouse-1, 10ul-mouse-1), and in gavage by 10% CMC (0.1ml-mouse-1), resvertrol (50.0mg·kg-1.d-1). Intratumoral inject by adenovirus every three days, a total of three times, and fed resveratrol every day for one month, two days after the last treatment all mice were sacrificed. The inhibitory rates and the sizes of the tumors were caculated. The expression levels of miR-181a miR-181a and SIRT 1 expression in different tissues were tested by Realtime-PCR and Western blot analysis respectively.Results:1. Compaired with normal endometrial tissue, miR-181a were up-regulated in EC samples, and were positively correlated with the degree of malignancy. SIRT 1 mRNA and protein expression were down-regulated. P-STAT 3 and AC-STAT 3 were up-regulated, STAT 3 was no significant change.2. The volume of xenografts reaching 50 mm3 typically needed 10-14 days after cells transplantation. The treatment with resveratrol and Ad-miR-181a had significant effects on tumor growth, compaired with A groups, the tumors of B group and F group were smaller significantly, C group were increases significantly, this difference reached statistical significance (p<0.001), E group were increases, this difference reached statistical significance (p= 0.023), the differences between E group and Agroup did not reach statistical significance (p= 0.252), as the differences between B group and F group (p= 0.195). The expression levels of miR-181a mRNA in B group was significantly lower than those in A group (p< 0.001), F group was lower(p= 0.001), C and D grooups were significantly higher, D and F groups were significantly lower than those in C and E groups (p< 0.001), and the differences between E groups and Agroup did not reach statistical significance(p= 0.092), as the differences between B group and F group (p= 0.076). The expression levels of SIRT 1 protein in B、D and F groups was significantly higher than those in A group, C group was significantly lower(p< 0.001), and the differences between E groups and Agroup did not reach statistical significance(p= 0.926), as the differences between B group and F group (p= 0.911).Conclusion: MiR-181a were up-regulated in EC samples than in normal endometrium, and were positively correlated with the degree of malignancy. SIRT 1 mRNA and protein expression were down-regulated. Overexpression miR-181a may silence the expression of SIRT 1 and could promote the growth of human endometrial carcinoma nude mouse xenografts markedly, and decrease the expression of SIRT 1. The resveratrol can inhibit the growth of tumor, and partially reversed the effects of Ad-miR-181a transfection on SIRT 1 protein expressions in vivo. It suggested that the anti-tumor mechanism of resveratrol might depend on miR-181a/SIRT1/STAT3 pathway in vivo. All the results suggest that miR-181a is an important dysregulated miRNAs in EC and may act as potential biomarker as well as a novel therapeutic target of EC, and provide new ideas for clinical treatment. |
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