Chen Wen-Ting, Yang Yu-Jia, Zhang Zhen-Dong, An Qiang, Li Na, Liu Wei, Yang Bing
Affiliated hospital of Zunyi Medical College, Zunyi, Guizhou, 563000, People's Republic of China.
J Ovarian Res. 2017 Jan 11;10(1):1. doi: 10.1186/s13048-016-0301-4.
We aimed to investigate the function of miR-1307 in chemoresistance and to explore its chemoresistance mechanism in ovarian cancer.
IC50 determination was used to test the chemoresistance profling in ovarian cancer cells. QRT-PCR or western blot was used to validate the expression level of miR-1307 and candidate gene or protein. Colony formation assay and FITC-labeled enhanced Annexin V immunofluorescence were used to compare cell proliferation and apoptosis ability, respectively. The potential target gene and its biological function of miRNA-1307 were also analyzed. Bioinformatics and Luciferase Reporter Gene Assay were conducted to validate the regulation of miRNA-1307 on the ING5 expression. Xenografts assay was used to demonstrate the inhibiting effect of miR-1307 ASO and Taxol therapy against ovarian cancer in vivo.
MiR-1307 was over-expressed in chemoresistant ovarian cancer cell line A2780/Taxol, and over-expression or loss of miR-1307 promoted or inhabited chemoresistance. And we also found that the over-expression of miR-1307 promoted proliferation and inhibited apoptosis in ovarian cancer cells. Besides, we demonstrated that ING5 was a direct target of miR-1307 and miR-1307 down-regulated the ING5 expression in ovarian cancer cells. Additionally, we showed that ING5 inhibited cell proliferation, promoted cell apoptosis and inhabited chemoresistance reversely. Furthermore, the up-regulated ability of cell apoptosis and down-regulated ability of chemoresistance following the loss of miR-1307 was reversed by adding ING5 siRNA in vitro. Finally, we proved the inhibiting effect of miR-1307 ASO and Taxol therapy by increasing the ING5 expression against ovarian cancer through xenografts assay in vivo.
Our results suggested that miR-1307 could promote ovarian cancer chemoresistance by targeting the ING5 expression and miR-1307 might serve as a therapeutic target for ovarian cancer.
我们旨在研究miR-1307在化疗耐药中的作用，并探索其在卵巢癌中的化疗耐药机制。
采用IC50测定法检测卵巢癌细胞的化疗耐药谱。运用QRT-PCR或蛋白质免疫印迹法验证miR-1307及候选基因或蛋白的表达水平。分别采用集落形成试验和FITC标记的膜联蛋白V增强免疫荧光法比较细胞增殖和凋亡能力。还分析了miRNA-1307的潜在靶基因及其生物学功能。进行生物信息学分析和荧光素酶报告基因检测以验证miR-1307对ING5表达的调控作用。采用异种移植试验在体内证明miR-1307反义寡核苷酸（ASO）和紫杉醇治疗对卵巢癌的抑制作用。
miR-1307在化疗耐药的卵巢癌细胞系A2780/Taxol中过表达，miR-1307的过表达或缺失分别促进或抑制化疗耐药。并且我们还发现miR-1307的过表达促进卵巢癌细胞增殖并抑制其凋亡。此外，我们证明ING5是miR-1307的直接靶标，且miR-1:307下调卵巢癌细胞中ING5的表达。另外，我们表明ING5抑制细胞增殖，促进细胞凋亡并反向抑制化疗耐药。此外，在体外通过添加ING5小干扰RNA（siRNA）可逆转miR-1307缺失后细胞凋亡上调能力和化疗耐药下调能力。最后，我们通过体内异种移植试验证明miR-1307 ASO和紫杉醇治疗通过增加ING5表达对卵巢癌具有抑制作用。
我们的结果表明，miR-1307可通过靶向ING5表达促进卵巢癌化疗耐药，且miR-1307可能成为卵巢癌的治疗靶点。
