Bioinformatics Identification of ZNFs/LINC00520/miR-181d/BCL2 Axis as a Novel Network in Cisplatin-Resistant Lung Adenocarcinoma Cells

Background: Resistance to cisplatin (DDP) leads to poor prognosis in patients with Lung Adenocarcinoma (LUAD) and limits its clinical application. It has been confirmed that autophagy promotes chemoresistance and, therefore, novel strategies to reverse chemoresistance by regulating autophagy are desperately needed. Methods: The differentially expressed lncRNAs (DElncRNAs), miRNAs (DEmiRNAs), and mRNAs (DEmRNAs) between A549 and A549/DDP cell lines were identified using the limma package in R, after gene expression profiles were obtained from Gene Expression Omnibus (GEO) database. By combining Autophagy-Related Genes (ARGs) from Human Autophagy Database (HADb), the interactions lncRNA-miRNAs and the interactions miRNAs-mRNAs respectively predicted by miRcode and miRDB/Targetscan database, the autophagy-related ceRNA network was constructed. Then, extraction of ceRNA subnetwork and Cox regression analyses were performed. A prognosis-related ceRNA subnetwork was constructed, and the upstream Transcription Factors (TFs) regulating lncRNAs were predicted by the JASPAR database. Finally, the expression patterns of candidate genes were further verified by quantitative real-time polymerase chain reaction (qRT-PCR) experiments. Results: A total of 3179 DEmRNAs, 180 DEmiRNAs, and 160 DElncRNAs were identified, and 35 DEmRNAs were contained in the HADb. Based on the ceRNA hypothesis, we established a ceRNA network, including 10 autophagy-related DEmRNAs, 9 DEmiRNAs, and 14 DElncRNAs. Then, LINC00520, miR-181d, and BCL2 were identified to construct a risk score model, which was confirmed to be a well-predicting prognostic factor. Furthermore, 5 TF ZNF family members were predicted to regulate LINC00520, whereas the RT-PCR results showed that the 5 ZNFs were consistent with the bioinformatics analysis. Finally, a ZNF regulatory LINC00520/miR-181d/BCL2 ceRNA subnetwork was constructed. Conclusions: An ZNFs/LINC00520/miR-181d/BCL2 axis as a novel network in DDP-resistant LUAD has been constructed successfully, which may provide potential therapeutic targets for LUAD.

针刺相对穴对穴区肥大细胞及其释放的组织胺和缓激肽的影响（英文）

目的:观察针刺大鼠相对穴阳陵泉与阴陵泉后穴区局部肥大细胞数量、脱颗粒情况及血清中组织胺、缓激肽的含量变化,探索"相对穴"协同增效的作用机制。方法:将40只Wistar大鼠采用随机数字表法随机分成空白对照组(K组)、针刺阳陵泉组(A组)、针刺阴陵泉组(B组)、针刺阳陵泉与阴陵泉组(AB组)及针刺阳陵泉与阴陵泉旁开组(CD组),每组8只。A组针刺阳陵泉;B组针刺阴陵泉;AB组针刺阳陵泉和阴陵泉;CD组分别采用阳陵泉和阴陵泉外侧旁开3 mm处作为旁开对照点,进行针刺。均选取双侧穴位。K组实验开始后采用与其余4组相同的方法固定,不给予针刺刺激。选用直径0.35 mm,长40 mm毫针,针刺得气后针柄接G6805-Ⅱ型电针治疗仪,选用疏密波,频率2 Hz/100 Hz,电流强度1 mA,以针柄轻微颤动且大鼠能保持安静为宜。每次电针持续刺激20 min。在开始后的第1、3、5、7天进行实验干预,共4次。各组大鼠于第7天干预完成2 h后取材。采用酶联免疫吸附法(ELISA)检测血清中组织胺和缓激肽的含量变化;穴区局部组织做冰冻切片,甲苯胺蓝染色,观察穴区肥大细胞(MCs)的变化。结果:与K组相比,A组、B组、AB组及CD组的MC数量及脱颗粒率均明显升高(均P<0.05),其中AB组的MC数量及脱颗粒率明显高于A组、B组和CD组(均P<0.05);组织胺含量由高到低的顺序为AB组>B组>A组>CD组>K组,且差异有统计学意义(均P<0.05);AB组、A组和B组的缓激肽含量明显高于K组和CD组(均P<0.05),A组和B组缓激肽含量差异,CD组与K组缓激肽含量差异均无统计学意义(均P>0.05)。结论:针刺"相对穴"阳陵泉与阴陵泉后穴区局部MC数量及脱颗粒率显著增高,血清中组织胺和缓激肽的含量升高,可能是"相对穴"配伍协同增效的机制之一。