桥接整合因子1在上皮性卵巢癌组织和细胞中的表达及其临床意义

目的：探讨桥接整合因子1(BIN1)在上皮性卵巢癌(EOC)组织中的表达及其临床意义,以及BIN1对EOC细胞A2780增殖、迁移和侵袭的影响。方法：收集2017年7月至2018年1月河北医科大学第四医院手术切除的67例EOC患者的肿瘤组织及同期因其他妇科疾病手术切除的30例非肿瘤患者的卵巢组织(正常对照组)标本。用免疫组织化学染色法检测EOC组织和非肿瘤卵巢组织中BIN1蛋白的表达水平,χ²检验分析BIN1表达与患者临床病理特征之间的关联,Kaplan-Meier法分析BIN1表达与患者的无病生存期(DFS)和总生存期(OS)之间的关系。用qPCR和WB法检测EOC细胞SKOV3、A2780和人卵巢上皮细胞IOSE80中BIN1 m RNA和蛋白的表达水平。利用基因转染技术将BIN1质粒CMV-MCS-GFP-SV40-NeomycinBIN1和空载体质粒CMV-MCS-GFP-SV40-Neomycin分别转染到A2780细胞以构建过表达BIN1细胞及其对照,用qPCR和WB法分别检测转染细胞中BIN1 mRNA和蛋白的表达水平,CCK-8、划痕愈合和Tran...     

LINC01503通过miR-342-3p/IGF2R轴促进上皮性卵巢癌的进展

目的：探讨LINC01503 在上皮性卵巢癌（EOC）中的表达水平和生物学功能及其可能的作用机制。方法：收集2015年5月至2016 年5月间在河北医科大学第四医院妇瘤科手术切除并经病理学确诊的85 例EOC患者的肿瘤组织和输卵管组织。常规培养人EOC 细胞A2780、SKOV3、OVCAR3 和OV90 及正常人卵巢上皮细胞IOSE80，将si-LINC01503、si-NC 及miR-342-3p mimic、miR mimic NC分别转染至SKOV3和A2780 细胞，分别作为si-LINC01503 组、si-NC 组、miR-342-3p mimic 组和miR mimic NC组。qPCR 法检测EOC组织和细胞中LINC01503 的表达水平，Kaplan-Meier 法分析LINC01503 表达水平与患者生存的关系。双荧光素酶报告基因实验验证LINC01503/miR-342-3p/IGF2R轴相关分子间的靶向关系。平板克隆、划痕愈合和Transwell 实验分别检测敲低LINC01503 及转染miR-342-3p mimic 对A2780 和SKOV3细胞增殖、迁移和侵袭能力的影响。WB法检测EOC细胞中LINC01503/miR-342-3p 通路对IGF2R蛋白表达的影响。构建A2780 细胞裸鼠移植瘤模型，观察敲低LINC01503 对移植瘤生长的影响。结果：EOC组织和细胞中LINC01503 表达水平分别显著高于输卵管组织和IOSE80 细胞（均P<0.01），LINC01503高表达组患者术后PFS 和OS 均显著短于LINC01503 低表达组患者（均P<0.01）。敲低LINC01503、转染miR-342-3p mimic 均可抑制EOC 细胞的增殖、迁移和侵袭能力（均P<0.01）。敲低LINC01503 可下调IGF2R的表达（P<0.01），这一现象可通过转染miR-342-3p inhibitor 挽救。敲低LINC01503 可抑制A2780 细胞裸鼠移植瘤的生长（P<0.01）。结论：在EOC 组织和细胞中呈高表达的LINC01503，与患者的不良预后密切相关，LINC01503可能通过吸附miR-342-3p影响IGF2R表达进而促进EOC的进展。     

C17orf76-AS1在卵巢上皮癌中的表达及其对卵巢上皮癌细胞SKOV3生物学功能的影响

目的 检测C17orf76-AS1在正常卵巢上皮细胞与卵巢上皮癌细胞系中的表达情况并探讨C17orf76-AS1过表达对卵巢上皮癌细胞系SKOV3生物学功能的影响。方法 通过定量PCR（QPCR）方法检测C17orf76-AS1在在卵巢上皮细胞IOSE80及不同卵巢上皮癌细胞系（A2780、SKOV3、OVCAR3、HO-8910和HO-8910PM）中的表达。在pcDNA3.1基础上构建C17orf76-AS1过表达质粒并采用Lipo2000瞬时转染SKOV3细胞,QPCR检测SKOV3细胞中C17orf76-AS1的过表达效率;CCK-8实验检测C17orf76-AS1过表达对SKOV3细胞增殖的影响;Transwell法和划痕实验检测C17orf76-AS1过表达对SKOV3细胞侵袭迁移能力的影响。结果 与正常卵巢上皮细胞IOSE80比较,C17orf76-AS1在A2780、SKOV3、 OVCAR3、HO-8910及HO-8910PM细胞中均呈低表达,分别下降了0.36、0.30、0.46、0.32、0.31倍（P＜0.05）。在SKOV3细胞中过表达C17orf76-AS1,CCK-8结果显示C17orf76-AS1过表达48 h后,SKOV3的增殖能力显著下降了约1.5倍;Transwell结果显示SKOV3细胞迁移能力下降约1.3倍,划痕实验也同样证实过表达C17orf76-AS1后,SKOV3细胞愈合能力显著低于对照组（P＜0.05）。结论 C17orf76-AS1在卵巢上皮癌细胞中异常低表达,其过表达显著降低了卵巢癌细胞增殖和迁移能力。     

LncRNA ITGA9-AS1在卵巢癌中的表达及与顺铂化疗敏感性的关系

目的:观察长链非编码RNA（LncRNA） ITGA9-AS1在卵巢癌中的表达水平,探究其与顺铂化疗敏感性的关系。方法:选取2017年01月至2019年01月本院收治的64例卵巢癌患者,卵巢摘除手术中采集卵巢癌组织,另收集因卵巢囊肿需作手术切除但已证实无瘤细胞的正常卵巢组织标本,采用实时定量PCR（RT-qPCR）检测卵巢癌及正常卵巢组织、顺铂（DDP）耐药卵巢癌细胞SKOV3/DDP、卵巢癌细胞（OVCAR5、OVCAR8、SKOV3）及永生化卵巢上皮细胞（IOSE29、IOSE80）中LncRNA ITGA9-AS1表达水平。采用慢病毒介导LncRNA ITGA9-AS1转染并筛选稳定细胞株,MTT法检测过表达LncRNA ITGA9-AS1对SKOV3、SKOV3/DDP增殖的影响及对不同浓度（0、1、2、4、8、16 mg/L）DDP的敏感性。结果:与正常卵巢组织比较,卵巢癌组织中LncRNA ITGA9-AS1表达水平降低（P＜0.05）。与IOSE29、IOSE80细胞比较,OVCAR5、OVCAR8、SKOV3及SKOV3/DDP细胞中LncRNA ITGA9-AS1表达水平显著降低（P＜0.05）;与SKOV3细胞比较,SKOV3/DDP细胞中LncRNA ITGA9-AS1表达水平显著降低（P＜0.05）。与空白对照组及慢病毒对照组比较,慢病毒过表达组SKOV3、SKOV3/DDP细胞中LncRNA ITGA9-AS1表达水平升高（P＜0.05）,增殖率降低（P＜0.05）,细胞存活率呈DDP浓度依赖降低（P＜0.05）。结论:LncRNA ITGA9-AS1在卵巢癌组织及癌细胞中低表达,过表达LncRNA ITGA9-AS1可增加卵巢癌SKOV3及卵巢癌顺铂耐药细胞SKOV3/DDP的顺铂化疗敏感性。     

Sphingosine kinase 1 as a potential therapeutic target in epithelial ovarian cancer

Sphingosine kinase 1 (SK1) is over‐expressed in multiple types of human cancer. SK1 has growth‐promoting effects and has been proposed as a potential therapeutic target. We investigated the therapeutic effects of SK1 inhibition in epithelial ovarian carcinoma (EOC). SK1 siRNA or inhibitors were tested in EOC cell lines, including A2780, SKOV3ip1, A2780‐CP20, SKOV3‐TR, ES2 and RMG2. Cells were treated with SK inhibitor or FTY720, and cell proliferation, apoptosis, angiogenesis and invasion were examined by MTT, FACS, ELISA and wound‐healing assays, respectively. In vivo experiments were performed to test the effects of FTY720 on tumor growth in orthotopic mouse xenografts of EOC cell lines A2780 or SKOV3ip1 and a patient‐derived xenograft (PDX) model of clear cell ovarian carcinoma (CCC). Blocking SK1 with siRNA or inhibitors significantly reduced proliferation, angiogenesis and invasion, and increased apoptosis in chemosensitive (A2780 and SKOV3ip1) and chemoresistant (A2780‐CP20, SKOV3‐TR, ES2 and RMG2) EOC cells. SK1 inhibitors also decreased the intracellular enzymatic activity of SK1. Furthermore, FTY720 treatment significantly decreased the in vivo tumor weight in xenograft models of established cell lines (A2780 and SKOV3ip1) and a PDX model for CCC compared to control (p < 0.05). These results support therapeutic targeting of SK1 as a potential new strategy for EOC.     

人上皮性卵巢癌组织中PFTK1的表达及其对细胞迁移、侵袭及增殖能力的影响

目的:研究PFTAIRE蛋白激酶1（PFTAIRE protein kinase 1,PFTK1）在人上皮性卵巢癌（epithelial ovarian cancer,EOC）组织中的表达,以及PFTK1基因的沉默对人卵巢癌细胞迁移、侵袭及增殖能力的影响。方法: 随机选取我院2015年6月至2016年11月收治的25例EOC患者为研究对象,采用Western blot检测癌组织以及癌旁组织中PFTK1蛋白的表达情况。选取PFTK1高表达的人卵巢癌细胞株SKOV3为研究对象,转染PFTK1-siRNA后,采用流式细胞仪检测PFTK1基因的沉默对SKOV3细胞周期的影响,采用细胞迁移和侵袭实验（Transwell）检测SKOV3细胞迁移、侵袭能力的变化,采用平板克隆形成实验检测SKOV3细胞增殖能力的变化。结果:Western blot检测结果显示,EOC患者癌组织中的PFTK1蛋白表达量显著高于癌旁组织（P＜0.01）;流式细胞仪检测结果显示,PFTK1-siRNA组细胞出现明显的生长抑制,细胞多停滞在G0/G1期,与对照组相比差异显著（P＜0.01）;细胞迁移和侵袭实验（Transwell）结果显示,PFTK1-siRNA组细胞的迁移和侵袭能力与对照组比较显著下降,且差异均显著（P＜0.01）;平板克隆形成实验结果显示,PFTK1-siRNA组细胞增殖能力显著降低,与对照组相比差异显著（P＜0.01）。结论:PFTK1在人EOC癌组织中呈现高表达状态;PFTK1基因沉默后,EOC细胞株SKOV3的细胞迁移、侵袭及增殖能力均受到不同程度的抑制,这将为基因治疗人卵巢癌提供一定的理论依据。     

浆细胞瘤多样异位基因1 在卵巢癌组织中的表达及其对SKOV3 细胞恶性生物学行为的影响

目的：研究人浆细胞瘤多样异位基因1（plasma-cytoma variant translocation gene 1, PVT1）对卵巢癌SKOV3 细胞增殖、迁移和侵袭能力的影响。方法：选取2015年11 月至2017 年4 月郑州大学第三附属医院妇产科收治的卵巢癌患者32例。利用实时荧光定量PCR检测PVT1 在32例卵巢癌组织及癌旁组织的表达。通过CCK-8法、划痕法及Transwell法检测PVT1 对卵巢癌细胞增殖、迁移及侵袭能力的影响。结果：PVT1在SKOV3细胞和卵巢癌组织表达水平均明显升高（均P<0.01）;PVT1表达水平与卵巢癌者组织学分级、FIGO分期及淋巴结转移具有相关性（P<0.05或P<0.01）。转染PVT1 siRNA36、48 h 后,SKOV3细胞中PVT1 表达水平明显下降（P<0.05）;敲减PVT1的表达能够降低SKOV3细胞的增殖能力（P<0.05）,抑制SKOV3细胞增殖、迁移和侵袭能力（P<0.05或P<0.01）。结论：PVT1 在卵巢癌中高表达,抑制PVT1表达能够抑制卵巢癌SKOV3细胞增殖、迁移和侵袭能力。     

LncRNA RP11-79H23.3在膀胱癌细胞中的作用及其发生、发展的研究

背景与目的：虽然许多长链非编码RNA(long non-coding RNA，lncRNA)的异常表达与膀胱癌的发生有密切关系，但对于lncRNA RP11-79H23.3暂未见报道。该研究旨在探讨lncRNA RP11-79H23.3在膀胱癌EJ细胞中的作用及其发生、发展的机制。方法：采用微阵列方法对4对膀胱癌患者的癌和癌旁组织进行组学分析，随后用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction，RTFQ-PCR)检测膀胱癌组织、癌旁组织及正常人膀胱上皮细胞sv-HUC-1、膀胱癌EJ细胞中RP11-79H23.3的表达。通过转染pIRES2-RP11-79H23.3上调该基因后，采用细胞计数试剂盒(cell counting kit-8，CCK-8)和EdU的方法检测EJ细胞的增殖活性，通过Transwell小室和平板划痕实验分别检测EJ细胞的侵袭和迁移能力，应用流式细胞术、Hoechst33342及Tunel检测细胞凋亡，应用细胞免疫荧光检测PTEN在膀胱癌细胞中的定位，采用鬼笔环肽染色观察细胞骨架形成，应用蛋白［质］印迹法(Western blot)分析过表达RP11-79H23.3后EJ细胞中PI3K/AKT信号通路中相关蛋白的表达情况。结果：LncRNA RP11-79H23.3在膀胱癌组织和膀胱癌EJ细胞中表达下调(P ＜0.001，P＜0.01)，pIRES2-RP11-79H23.3转染EJ细胞结果显示，RP11-79H23.3的表达量较转染前显著增加(P＜0.01)。上调RP11-79H23.3的表达可诱导膀胱癌EJ细胞的凋亡，相反，转染pIRES2-EGFP可促进EJ细胞的增殖、侵袭和迁移能力，同时，Western blot结果显示，转染pIRES2-RP11-79H23.3后可上调PTEN在EJ细胞中的表达，下调p-PI3K、p-AKT及p-Gsk3β蛋白的表达(P＜0.05)。结论：LncRNA RP11-79H23.3在膀胱癌组织和膀胱癌EJ细胞中低表达(P＜0.001，P＜0.01)，并且过表达RP11-79H23.3会降低膀胱癌细胞增殖、侵袭和迁移能力，其作用机制可能与PI3K/AKT信号通路有关。提示lncRNA RP11-79H23.3在膀胱癌恶性肿瘤中发挥重要的作用，可能会成为治疗膀胱癌的药物作用靶点。     

LncRNA RP11-436H11.5, functioning as a competitive endogenous RNA,upregulates BCL-W expression by sponging miR-335-5p and promotes proliferation and invasion in renal cell carcinoma

Background

Accumulating evidence indicates that long non-coding RNAs (lncRNAs) play a crucial role in tumorigenesis. Here, we report a novel lncRNA, RP11-436H11.5, that regulates renal cell carcinoma (RCC) cell proliferation and invasion by sponging miR-335-5p.

Methods

Expression of lncRNA RP11-436H11.5 was determined by a qRT-PCR assay in RCC tissues. RCC cell proliferation and invasion were measured by a cell proliferation assay and a transwell invasion assay. Expression of BCL-W was detected by a western blot assay. Interactions between lncRNA RP11-436H11.5 and miR-335-5p were measured by a luciferase reporter assay and a RNA-pull down assay. In vivo experiments were used to detect tumor formation.

Results

In this study, the qRT-PCR results illustrated that lncRNA RP11-436H11.5 was more highly expressed in RCC tissues than in adjacent normal renal tissues. The results of survival analysis indicated that patients in the high lncRNA RP11-436H11.5 group presented significantly worse outcomes compared with those in the low lncRNA RP11-436H11.5 group. Downregulation of lncRNA RP11-436H11.5 suppressed RCC cell proliferation and invasion in vitro and in vivo. Luciferase reporter assay results demonstrated that lncRNA RP11-436H11.5 enhanced BCL-W expression by regulating miR-335-5p expression. LncRNA RP11-436H11.5 could function as a miR-335-5p decoy to derepress expression of BCL-W.

Conclusions

LncRNA RP11-436H11.5 could function as a competing endogenous RNA to promote RCC cell proliferation and invasion, which might serve as a therapeutic application to suppress RCC progression.

     

伽马-分泌酶抑制剂下调Notch1可诱导卵巢癌细胞A2780生长抑制及凋亡

目的 探讨γ-分泌酶抑制剂对Notch1的抑制作用及对卵巢癌细胞生长抑制及凋亡的影响。方法 采用Western blot及实时定量PCR检测四株卵巢癌细胞（A2780, SKOV3, HO-8910, HO-8910PM）及一株卵巢上皮细胞（IOSE144）Notch1及其下游基因hes1的表达情况;采用MTT、流式细胞术、ELISA及克隆形成实验检测γ-分泌酶抑制剂（DAPT）对卵巢癌细胞的影响。结果 Notch1、hes1在IOSE 144及四株卵巢癌细胞系中均有表达,且在A2780细胞系中的表达最高;DAPT下调Notch1可抑制A2780细胞生长、诱导细胞G1期阻滞、诱导细胞凋亡并呈现时间和剂量依赖性,同时A2780细胞中DAPT 下游hes1基因的表达也呈现时间和剂量依赖性。结论 γ-分泌酶抑制剂DAPT可抑制卵巢癌细胞A2780生长、诱导细胞凋亡,γ-分泌酶抑制剂下调Notch1可能是卵巢癌治疗的潜在靶点。     

Differential cellular responses induced by dorsomorphin and LDN‐193189 in chemotherapy‐sensitive and chemotherapy‐resistant human epithelial ovarian cancer cells

Inherent or acquired drug resistance is a major contributor to epithelial ovarian cancer (EOC) mortality. Novel drugs or drug combinations that produce EOC cell death or resensitize drug resistant cells to standard chemotherapy may improve patient treatment. After conducting drug tolerability studies for the multikinase inhibitors dorsomorphin (DM) and it is structural analogue LDN‐193189 (LDN), these drugs were tested in a mouse intraperitoneal xenograft model of EOC. DM significantly increased survival, whereas LDN showed a trend toward increased survival. In vitro experiments using cisplatin (CP)‐resistant EOC cell lines, A2780‐cp or SKOV3, we determined that pretreatment or cotreatment with DM or LDN resensitized cells to the killing effect of CP or carboplatin (CB). DM was capable of blocking EOC cell cycle and migration, whereas LDN produced a less pronounced effect on cell cycle and no effect on migration. Subsequent analyses using primary human EOC cell samples or additional established EOC cells lines showed that DM or LDN induced a dose‐dependent autophagic or cell death response, respectively. DM induced a characteristic morphological change with the appearance of numerous LC3B‐containing acidic vacuoles and an increase in LC3BII levels. This was coincident with a decrease in cell growth and the altered cell cycle consistent with DM‐induced cytostasis. By contrast, LDN produced a caspase 3‐independent, reactive oxygen species‐dependent cell death. Overall, DM and LDN possess drug characteristics suitable for adjuvant agents used to treat chemotherapy‐sensitive and ‐resistant EOC.     

Long non‐coding RNA urothelial cancer‐associated 1 promotes bladder cancer cell migration and invasion by way of the hsa‐miR‐145–ZEB1/2–FSCN1 pathway

Numerous studies suggest that several long non‐coding RNAs (lncRNAs) play critical roles in bladder cancer development and progression. Long non‐coding RNA urothelial cancer‐associated 1 (lncRNA‐UCA1) is highly expressed in bladder cancer tissues and cells, and it has been shown to play an important role in regulating aggressive phenotypes of bladder cancer cells. However, little is known about the molecular mechanism of lncRNA‐UCA1‐mediated bladder cancer cell migration and invasion. Here, we show that overexpression of lncRNA‐UCA1 could induce EMT and increase the migratory and invasive abilities of bladder cancer cells. Mechanistically, lncRNA‐UCA1 induced EMT of bladder cancer cells by upregulating the expression levels of zinc finger E‐box binding homeobox 1 and 2 (ZEB1 and ZEB2), and regulated bladder cancer cell migration and invasion by tumor suppressive hsa‐miR‐145 and its target gene the actin‐binding protein fascin homologue 1 (FSCN1). Furthermore, we also observed a positive correlation between lncRNA‐UCA1 and ZEB1/2 expression, and a negative correlation between lncRNA‐UCA1 and hsa‐miR‐145 expression in bladder cancer specimens. Importantly, we found that lncRNA‐UCA1 repressed hsa‐miR‐145 expression to upregulate ZEB1/2, whereas the suppression of hsa‐miR‐145 could upregulate lncRNA‐UCA1 expression in bladder cancer cells. Moreover, the binding site for hsa‐miR‐145 within exons 2 and 3 of lncRNA‐UCA1 contributed to the reciprocal negative regulation of lncRNA‐UCA1 and hsa‐miR‐145. Taken together, our results identified that lncRNA‐UCA1 enhances bladder cancer cell migration and invasion in part through the hsa‐miR‐145/ZEB1/2/FSCN1 pathway. Therefore, lncRNA‐UCA1 might act as a promising therapeutic target for the invasion and metastasis of bladder cancer.     

Exosomal transfer of miR-429 confers chemoresistance in epithelial ovarian cancer

The development of multidrug resistance during chemotherapy is the main obstacle for epithelial ovarian cancer (EOC) treatment. Exosomal transfer of carcinogenic microRNAs (miRNAs) might strengthen chemoresistance in recipient cells. Here, we identified through microarray analysis higher miR-429 expression in multidrug-resistant SKOV3 cells and their secreted exosomes (SKOV3-EXO) than in sensitive A2780 cells and their secreted exosomes. SKOV3-derived exosomes were internalized by A2780 cells, which permitted the transfer of miR-429. Exosomal miR-429 enhanced the proliferation and drug resistance of A2780 cells by targeting calcium-sensing receptor (CASR)/STAT3 pathway in vitro and in vivo. In addition, NF-κB-p65 was predicted to bind to the miR-429 promoter region, and the inhibition of NF-κB reduced the expression of miR-429 and led to the sensitivity of EOC cells. Consistently, A2780 cells co-incubated with SKOV3 pretreated with an NF-κB inhibitor or miR-429 antagomir showed sensitivity to cisplatin and exhibited attenuated cell proliferation. Based on our data, exosomal miR-429 functions as a primary regulator of the chemoresistance and malignant phenotypes of EOC by targeting CASR through a mechanism promoted by NF-κB and might be a therapeutic target for EOC.     

lncRNA HEIG对宫颈癌细胞增殖、凋亡和迁移侵袭的影响及其分子机制

目的:探讨长链非编码RNA(lncRNA)HEIG在宫颈癌组织和细胞中的表达及其生物学功能。方法:采用实时荧光定量PCR检测患者癌组织和癌旁组织、正常宫颈上皮细胞HCerEpic、宫颈癌细胞Hela和C33A中lncRNA HEIG的表达;敲低Hela细胞中HEIG表达,MTT法和流式细胞术分别检测Hela细胞的增殖、凋亡情况;Transwell小室法检测Hela细胞的迁移侵袭能力;Western blot检测AKT信号通路的变化。结果:lncRNA HEIG在宫颈癌组织和细胞中高表达;敲低HEIG可显著抑制宫颈癌细胞Hela的细胞增殖,促进细胞凋亡,抑制细胞迁移和侵袭能力,同时抑制AKT信号通路的活化。结论:lncRNA HEIG有可能作用于AKT信号通路抑制宫颈癌细胞增殖、促进细胞凋亡、抑制细胞迁移侵袭。     

miR-204在卵巢癌中的表达及对卵巢癌细胞增殖、侵袭的影响

目的:探讨miR-204 在卵巢癌组织中的表达情况及其对卵巢癌细胞增殖和侵袭能力的影响.方法:RT-PCR法检测卵巢癌细胞 SKOV-3及卵巢正常上皮细胞IOSE80中 miR-204 的表达水平;CCK-8法检测侵染后SKOV-3及IOSE8细胞的增殖;流式细胞仪检测各组细胞的凋亡;Transwell 法检测miR-204转染后SKOV-3 细胞的侵袭能力.结果:SKOV-3内miR-204表达较IOSE80明显降低,二者之间具有显著性差异(P=0.008 2,P<0.05);感染后各组细胞内miR-204表达水平及细胞增殖能力、细胞凋亡能力及侵袭能力方面,实验组与空白组和对照组之间都有显著性差异(P<0.05);而空白对照组与载体对照组之间都没有显著性差异(P>0.05),说明过表达miR-204能够抑制细胞的增殖能力,促进细胞凋亡并且影响细胞侵袭能力.结论:过表达miR-204能够抑制细胞的增殖能力,促进细胞凋亡并且影响细胞侵袭能力.     

长链非编码RNA RP11-385J1.2通过靶向微小RNA-370-3p调控甲状腺癌细胞增殖、侵袭及凋亡的作用机制

目的探讨长链非编码RNA(lncRNA)RP11-385J1.2对甲状腺癌细胞增殖、侵袭和凋亡的影响。方法采用实时荧光聚合酶链反应(PCR)检测人甲状腺癌细胞株SW579、FTC-133、TPC-1、K1和正常甲状腺细胞Nthyori 3-1中RP11-385J1.2和微小RNA(miRNA)-370-3p的表达情况。采用抑制剂si-RP11-385J1.2和(或)anti-miRNA-370-3p转染SW579细胞,噻唑蓝(MTT)法检测细胞增殖,Transwell实验检测细胞侵袭,流式细胞术检测细胞凋亡,蛋白质印记法(Western blot)检测凋亡相关蛋白B细胞淋巴瘤/白血病-2(Bcl-2)和Bax的表达情况,双荧光素酶报告系统验证RP11-385J1.2和miRNA-370-3p的靶向关系。结果人甲状腺癌细胞株SW579、FTC-133、TPC-1、K1中RP11-385J1.2的表达水平均高于正常甲状腺细胞Nthy-ori 3-1,miRNA-370-3p的表达水平均低于正常甲状腺细胞Nthy-ori 3-1,差异均有统计学意义(P﹤0.05)。敲减RP11-385J1.2可抑制SW579细胞增殖和侵袭并促进细胞凋亡,RP11-385J1.2能够靶向负调控miRNA-370-3p的表达,下调miRNA-370-3p的表达可逆转敲减RP11-385J1.2对SW579细胞增殖、侵袭和凋亡的影响。结论lncRNA RP11-385J1.2通过靶向下调miRNA-370-3p的表达促进甲状腺癌SW579细胞增殖和侵袭并抑制细胞凋亡。RP11-385J1.2和miRNA-370-3p是甲状腺癌的潜在分子靶点。