目的：探讨LINC00462 招募转录因子 MYC 激活 ABCC3 对肾透明细胞癌（ccRCC）顺铂敏感性的影响及其机制。方法：数据库分析ccRCC 组织中ABCC3、MYC和LINC00462 的表达及其相关性，并分析ABCC3 基因的富集通路。常规培养人肾小管上皮细胞（HK-2）和ccRCC 细胞（A-498、786-O 和 Caki-2），将 si-LINC00462、oe-ABCC3、si-ABCC3、si-MYC、si-LINC00462-NC、oe-ABCC3-NC、si-ABCC3-NC 和 si-MYC-NC 核酸序列分别转染 A-498 或786-O 细胞，分为si-LINC00462组、si-LINC00462-NC 组、oe-ABCC3 组、oe-ABCC3-NC 组、si-ABCC3 组、si-ABCC3-NC 组、si-MYC 组、si-MYC-NC 组；用2-脱氧-D-葡萄糖（2-DG）进行回复实验，构建oe-NC+PBS 组、oe-ABCC3+PBS组、oe-ABCC3+2-DG组；为探究ccRCC细胞LINC00462/MYC/ABCC3 轴对顺铂敏感性的影响，构建si-NC+oe-NC 组、si-LINC00462+oe-NC 组、si-LINC00462+oe-ABCC3 组。qPCR 法检测ABCC3、MYC和LINC00462 在ccRCC细胞中的表达，CCK-8法检测细胞增殖活力，CCK-8法分析梯度浓度顺铂处理ccRCC细胞后IC50值，WB法检测糖酵解代谢途径相关蛋白的表达，Seahorse XP96 法检测各处理组细胞的胞外酸化率（ECAR）和耗氧率（OCR），试剂盒检测细胞中丙酮酸、乳酸、ATP水平。双荧光素酶报告基因和染色质免疫共沉淀（ChIP）实验验证ABCC3与MYC间的结合关系，RNA结合蛋白免疫沉淀（RIP）实验验证LINC00462 和MYC的结合关系。结果：数据库分析和qPCR实验结果显示，ABCC3在ccRCC组织和细胞中呈高表达，差异基因富集在糖酵解通路上。敲减或过表达ABCC3能够增加A-498细胞或降低786-O细胞对顺铂的敏感性，ABCC3可通过促进有氧糖酵解抑制A-498 细胞对顺铂的敏感性，2-DG处理可以逆转过表达ABCC3对ccRCC 细胞对顺铂敏感性的抑制作用。MYC可直接和ABCC3结合，LINC00462 可招募转录因子MYC；敲低LINC00462 可抑制ABCC3的表达，敲低LINC00462 可抑制ccRCC细胞的有氧糖酵解，并提高其对顺铂敏感性；而进一步过表达ABCC3可逆转敲低LINC00462 对ccRCC 细胞有氧糖酵解的抑制作用和顺铂敏感性的提高。结论：LINC00462 通过招募转录因子MYC 激活ABCC3的表达促进ccRCC细胞的糖酵解，进而促进ccRCC细胞对顺铂敏感性。

Objective：：To investigate the effect of LINC00462 binding to transcription factor MYC to activate ABCC3 on cisplatin sensitivity in clear cell renal cell carcinoma (ccRCC) and its potential mechanisms. Methods: The expression of ABCC3, MYC and LINC00462 and their correlation in ccRCC were analyzed, and the enrichment pathway of ABCC3 was also analyzed. Human renal tubular epithelial cells (HK-2) and ccRCC cells (A-498, 786-O and Caki-2) were conventionally cultured. Nucleic acid sequences of si-LINC00462,si-LINC00462-NC, oe-ABCC3, oe-ABCC3-NC, si-ABCC3, si-ABCC3-NC, si-MYC and si-MYC-NC were transfected into A-498 or 786-O cells respectively, namely si-LINC00462 group, si-LINC00462-NC group, oe-ABCC3 group, oe-ABCC3-NC group, si-ABCC3 group,si-ABCC3-NC group, si-MYC group and si-MYC-NC group. 2-deoxy-D-glucose (2-DG) was used in the rescue experiment, and oe-NC+PBS group, oe-ABCC3+PBS group and oe-ABCC3+2-DG group were established. To investigate the effects of LINC00462/MYC/ABCC3 axis on the cisplatin sensitivity of ccRCC cells, si-NC+oe-NC group, si-LINC00462+oe-NC gorup, and si-LINC00462+oe-ABCC3 group were constructed. The expression of ABCC3, MYC and LINC00462 in ccRCC cells was detected by qPCR. CCK-8 method was used to measure cell viability and the IC50 of cisplatin inhibiting the proliferation of ccRCC cells. WB was used to detect the expression of glycolytic metabolism-related proteins. Seahorse XP96 was used to analyze the extracellular acidification rate (ECAR) and oxygen consumption rate(OCR) in the cells of different treatment groups. The levels of pyruvate, lactic acid and ATP in the cells were detected by corresponding kits.The binding relationship between ABCC3 and MYC was verified by double luciferase reporter gene assay and chromatin immunoprecipitation (ChIP) assay, and the binding relationship between LINC00462 and MYC was verified using RNA-binding protein immunoprecipitation (RIP) experiment. Results: Database analysis and qPCR experiments showed that ABCC3 was highly expressed in ccRCC tissues and cells and was enriched in the glycolytic pathway. Knockdown of ABCC3 could promoted the sensitivity of A-498 cells to cisplatin, while overexpression of ABCC3 could reduce the sensitivity of 786-O cells to cisplatin. ABCC3 could inhibit the sensitivity of A-498 cells to cisplatin through promoting glycolysis. 2-DG treatment reversed the inhibitory effect of ABCC3 overexpression on cisplatin sensitivity of ccRCC cells. MYC could directly bind to ABCC3, and LINC00462 could recruit MYC. Knockdown of LINC00462 inhibited the expression of ABCC3, suppressed aerobic glycolysis of ccRCC cells and increased their sensitivity to cisplatin; however, further over-expression of ABCC3 reversed the effects of LINC00462 knockdown on inhibiting aerobic glycolysis and promoting cisplatin sensitivity in ccRCC cells. Conclusion：： LINC00462 promotes glycolysis of ccRCC cells and further inhibits their sensitivity to cisplatin by recruiting the transcription factor MYC to activate ABCC3 expression.

河北省医学科学研究课题（No. 20220452）