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OBJECTIVE: To investigate the effect of shRNA targeting HMGB1 on HMGB1 expression and its relationship with invasion and migration of endometrial carcinoma cells. METHODS: HMGB1 short hairpin RNA (pshRNA-1 / HMGB1, pshRNA-2 / HMGB1, pshRNA-3 / HMGB1) was constructed by RNA interference technique, and negative control group (HMGB1 / p-NC ) And liposome transfection group (Lipo group) were transfected into human endometrial carcinoma cells (HEC-1A) by lipofectamine method. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting The expression of HMGB1 in the HEC-1A cells transfected with HMGB1 shRNA was observed by Transwell chamber method. Cell scratches were observed in the cells of HEC-1A cells after transfection with HMGB1 shRNA. Migration situation. RESULTS: RT-PCR showed that the relative expression levels of HMGB1 mRNA were 0.192 0.006, 0.055 0.002 and 0.123 0.086, respectively. Compared with Lipo group (0.268, 0.008) and HMGB1 / p- NC group (0.270, 0.004) was significantly decreased (P <0.05), the maximum inhibition rate was 28.4%, 79.5% and 54.1% respectively. Western blot showed that the relative expression levels of HMGB1 protein were 0.259 ,0.129, 0.032 ,0.002 and 0.104 ,0.007, respectively (P <0.05) compared with Lipo group (0.347 ,0.007) and HMGB1 / p-NC group (0.349 ,0.007) , The maximum inhibition rate was 25.4%, 90.8% and 70.0% respectively. The number of cells passing through the Transwell chamber was significantly lower than that in the HMGB1 / p-NC and Lipo groups (P <0.05) at 48 h after inoculation. Sclerated healing experiments also showed that the sclerosis healing rate of the experimental group was significantly lower than that of HMGB1 / p-NC group and Lipo group (P <0.05). CONCLUSION: ShRNA targeting HMGB1 can inhibit the expression of HMGB1, and the invasion and migration ability of endometrial carcinoma cells is significantly decreased after transfection.
Luan X, Ma C, Wang P, Lou F. HMGB1 is negatively correlated with the development of endometrial carcinoma and prevents cancer cell invasion and metastasis by inhibiting the process of epithelial-to-mesenchymal transition. OncoTargets and therapy. 2017;10:1389-1402. doi:10.2147/OTT.S123085.
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