ABSTRACT

Purpose

Chemoresistance remains the primary obstacle threatening the prognosis of retinoblastoma (RB). microRNAs (miRNAs) are acknowledged as critical regulators of drug resistance. This study explored the molecular mechanism of miR-130a-3p affecting the chemosensitivity of RB to vincristine (VCR).

Methods

miR-130a-3p expression of human retinal astrocytes and RB cell lines (Y79, WERI-Rb-1, SO-Rb50, and SO-Rb70) was detected using RT-qPCR. VCR-resistant RB cell line Y79/VCR was induced. miR-130a-3p expression of Y79/VCR cell line and its corresponding parental cell line was detected. Y79/VCR cells were subjected to miR-130a-3p overexpression treatment. The cell proliferation was measured using MTT assay, and the IC50 value and drug resistance index were examined using CCK-8 assay. The targeting relationship between miR-130a-3p and PAX6 was predicted through bioinformatics analysis and verified using dual-luciferase assay. Functional rescue experiments were conducted to confirm the role of PAX6 in chemosensitivity of RB cells. The effect of miR-130a-3p on tumorigenesis and VCR sensitivity was observed in vivo.

Results

miR-130a-3p was downregulated in VCR-resistant RB cells. Overexpression of miR-130a-3p repressed the proliferation of Y79/VCR cells and enhanced chemosensitivity. miR-130a-3p targeted PAX6 expression. Overexpression of PAX6 reversed the effect of miR-130a-3p on chemosensitivity of Y79/VCR cells. Overexpression of miR-130a-3p suppressed tumor growth and reduced VCR resistance in vivo.

Conclusions

miR-130a-3p enhanced the chemosensitivity of Y79 RB cells to VCR by targeting PAX6 expression.

中文翻译：

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miR-130a-3p 通过靶向 PAX6 表达增强 Y79 视网膜母细胞瘤细胞对长春新碱的化学敏感性

摘要

目的

化疗耐药仍然是威胁视网膜母细胞瘤 (RB) 预后的主要障碍。microRNAs (miRNAs) 被认为是耐药性的关键调节因子。本研究探讨了miR-130a-3p影响RB对长春新碱（VCR）化学敏感性的分子机制。

方法

使用 RT-qPCR 检测人视网膜星形胶质细胞和 RB 细胞系（Y79、WERI-Rb-1、SO-Rb50 和 SO-Rb70）的 miR-130a-3p 表达。诱导了VCR抗性RB细胞系Y79/VCR。检测Y79/VCR细胞系及其对应的亲代细胞系的miR-130a-3p表达。对 Y79/VCR 细胞进行 miR-130a-3p 过表达处理。MTT法检测细胞增殖情况，CCK-8法检测IC50值和耐药指数。通过生物信息学分析预测 miR-130a-3p 和 PAX6 之间的靶向关系，并使用双荧光素酶测定进行验证。进行了功能性拯救实验以确认 PAX6 在 RB 细胞化学敏感性中的作用。在体内观察到 miR-130a-3p 对肿瘤发生和 VCR 敏感性的影响。

结果

miR-130a-3p 在 VCR 抗性 RB 细胞中下调。miR-130a-3p 的过表达抑制了 Y79/VCR 细胞的增殖并增强了化学敏感性。miR-130a-3p 靶向 PAX6 表达。PAX6 的过表达逆转了 miR-130a-3p 对 Y79/VCR 细胞化学敏感性的影响。miR-130a-3p 的过表达抑制了肿瘤生长并降低了体内VCR 抗性。

结论

miR-130a-3p 通过靶向 PAX6 表达增强了 Y79 RB 细胞对 VCR 的化学敏感性。