lncRNA SNHG12与ELAVL1相互作用激活PI3K/AKT信号通路促进前列腺癌细胞多西他赛的耐药机制

doi:10.12122/j.issn.1673-4254.2026.01.20

摘要/Abstract

摘要：

目的探讨lncRNA SNHG12对前列腺癌（PCa）多西他赛（DTX）耐药的影响及潜在机制。方法使用梯度DTX处理PC-3细胞获得DTX耐药的PC-3细胞（PC-3R），通过在雄性BALB/c裸鼠左背部注射PC-3R细胞构建PCa荷瘤模型。动物实验分为对照组（NC），DTX组，sh-SNHG12组，DTX+sh-SNHG12组， 5只/组。通过RT-qPCR、Western blotting、免疫荧光、免疫组化检测关键基因和蛋白的表达，通过CCK-8、克隆形成以及Transwell迁移实验评估细胞的增殖情况，通过RIP-qPCR检测RNA与蛋白之间的结合。结果 SNHG12在PC-3R细胞中表达上调（P<0.001），敲低SNHG12可抑制PC-3R细胞的增殖和细胞迁移能力以及裸鼠荷瘤组织生长（P<0.001），10 nmol/L DTX处理对PC-3R细胞增殖及迁移没有显著影响，而在DTX处理的基础上敲低SNHG12可抑制PC-3R细胞增殖、迁移以及裸鼠荷瘤组织的生长（P<0.001）。在PC-3R细胞中ELAVL1的表达上调（P<0.001），PC-3R细胞及荷瘤组织中PI3K/AKT信号通路活化水平也上调，并且额外PI3K激活剂740 Y-P处理可削弱敲低SNHG12的作用。PC-3R细胞中的SNHG12可与ELAVL1结合。机制研究发现，SNHG12通过与ELAVL1结合来激活PI3K/AKT信号通路，进而导致PCa DTX耐药。结论敲低SNHG12可以抑制PCa DTX耐药，为PCa DTX增敏疗法的开发提供了潜在干预靶点。

关键词: 前列腺癌, 耐药, 多西他赛, lncRNA SNHG12, ELAVL1, PI3K/AKT信号通路

Abstract:

Objective To investigate the regulatory role of lncRNA SNHG12 in docetaxel (DTX) resistance of prostate cancer (PCa) cells. Methods Tumor-bearing male BALB/c nude mouse models were stablished by dorsal subcutaneous injection of PC-3 cells or DTX-resistant PC-3 (PC-3R) cells, either with or without transfection with sh-SNHG12 prior to the injection (n=5). The expressions of the key genes and proteins in the tumor tissues were detected using RT-qPCR, Western blotting, immunofluorescence staining or immunohistochemistry. The proliferation and migration of the treated cells were evaluated with CCK-8, clone formation and Transwell migration assays. RIP-qPCR technique was used to determine the binding between the RNAs and proteins. Results SNHG12 expression was significantly up-regulated in PC-3R cells. SNHG12 knockdown effectively inhibited proliferation and migration of PC-3R cells in vitro and suppressed tumor growth in nude mice. While 10 nmol/L DTX treatment alone did not significantly affect proliferation or migration of PC-3R cells, its combination with SNHG12 knockdown strongly inhibited cell proliferation and migration both in vitro and in the tumor-bearing mice. The expression of ELAVL1 was obviously up-regulated in PC-3R cells, and increased activation level of PI3K/AKT signaling pathway was detected in both PC-3R cells and the xenografts. The effect of SNHG12 knockdown was significantly weakened by treatment with the PI3K activator 740 Y-P. SNHG12 was found to bind to ELAVL1 in PC-3R cells, and mechanistic studies showed that their binding activated the PI3K/AKT signaling pathway to result in DTX resistance in PCa. Conclusion SNHG12 knockdown inhibits DTX resistance of PCa cells by reducing SNHG12 binding to ELAVL1 to inhibit the activation the PI3K/AKT signaling pathway.

Key words: prostate cancer, drug resistance, docetaxel, lncRNA SNHG12, ELAVL1, PI3K / AKT signaling pathway

赵铖, 李稳, 郑宝寿, 王光明, 肖芝松, 李云鹏. lncRNA SNHG12与ELAVL1相互作用激活PI3K/AKT信号通路促进前列腺癌细胞多西他赛的耐药机制[J]. 南方医科大学学报, 2026, 46(1): 183-190.

Cheng ZHAO, Wen LI, Baoshou ZHENG, Guangming WANG, Zhisong XIAO, Yunpeng LI. Overexpression of lncRNA SNHG12 promotes docetaxel resistance of prostate cancer cells by activating PI3K/AKT signaling via interacting with ELAVL1[J]. Journal of Southern Medical University, 2026, 46(1): 183-190.

图/表 6

表1 引物序列信息列表

Tab.1 Primer sequences for RT-qPCR

Genes	Primer (5'-3′)
SNHG12	F:ATGAAATGCAGGGGACCTGG
SNHG12	R:TGTAACATGAATCTTAAAGCACAGC
ELAVL1	F:AACTACGTGACCGCGAAGG R:CGCCCAAACCGAGAGAACA
β-actin	F:CATGTACGTTGCTATCCAGGC
β-actin	R:CTCCTTAATGTCACGCACGAT

图1 多西他赛耐药细胞的构建以及SNHG12的表达鉴定

Fig.1 Construction of docetaxel-resistant PC-3 cells and identification of SNHG12 expression. A: CCK-8 assay for detecting cell viability. B: RT-qPCR for detecting expression of SNHG12 in PC-3 and PC-3R cells. n=3, ***P<0.001 vs PC-3 group.

图2 敲低SNHG12抑制前列腺癌细胞的多西他赛耐药

Fig.2 Knockdown of SNHG12 inhibits docetaxel resistance in prostate cancer cells. A: Transfection efficiency of sh-SNHG12 detected by RT-qPCR. B: CCK-8 assay for assessing cell viability. C: Clone formation assay for assessing cell clone formation ability. D: Transwell assay for assessing cell migration ability (Scale bar=100 μm). n=3, **P<0.001, ***P<0.001 vs NC group; ###P<0.001 vs DTX group.

图3 敲低SNHG12通过PI3K/AKT信号通路抑制前列腺癌细胞的多西他赛耐药

Fig.3 Knockdown of SNHG12 inhibits docetaxel resistance in prostate cancer cells through the PI3K/AKT signaling pathway. A: Western blotting for detecting expressions of p-PI3K/PI3K and p-AKT/AKT in PC-3 and PC-3R cells. B: CCK-8 assay for assessing viability of PC-3R cells. C: Clone formation assay for assessing clone formation ability of PC-3R cells. D: Transwell assay for assessing migration ability of PC-3R cells (Scale bar=100 μm). n=3, ***P<0.001 vs PC-3 group/DTX group; ##P<0.01, ###P<0.001 vs DTX+sh-SNHG12 group.

图4 SNHG12与ELAVL1相互作用激活PI3K/AKT信号通路

Fig.4 SNHG12 interacts with ELAVL1 to activate the PI3K/AKT signaling pathway. A: Immunofluorescence staining for detecting ELAVL1 expression in PC-3 and PC-3R cells (Scale bar=10 μm). B: RIP-qPCR for detecting enrichment level of SNHG12 on ELAVL1 in PC-3 and PC-3R cells. C: RT-qPCR for detecting transfection efficiency of sh-ELAVL1. D: Western blotting for detecting expressions of p-PI3K/PI3K and p-AKT/AKT in PC-3R cells. n=3, **P<0.01, ***P<0.001 vs PC-3 group/IgG group/NC group.

图5 敲低SNHG12抑制前列腺癌多西他赛耐药

Fig.5 Knockdown of SNHG12 inhibits docetaxel resistance in prostate cancer. A: Gross observation of the dissected tumors from the nude mouse models. B: Tumor volume changes in each group. C: Tumor weight changes in each group. D: Immunohistochemical detection of Ki67 expression in the tumor tissues in each group (Scale bar=25 μm). E: Western blotting for detecting expressions of p-PI3K/PI3K and p-AKT/AKT in the tumor tissues. n=5, *P<0.05, ***P<0.001 vs NC group; ###P<0.001 vs DTX group.

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