下调Beclin1 抑制卵巢癌A2780/DDP细胞对顺铂的耐药性

目的：探究敲减Beclin1 表达对卵巢癌细胞A2780 对顺铂耐药的影响及其相关机制。方法：以Western blotting 及qPCR 检测卵巢癌细胞株A2780 及耐药细胞株A2780/DDP 中Beclin1 的表达情况；A2780/DDP 细胞转染Beclin1 siRNA 后，用MTT法检测细胞对顺铂敏感性的变化，克隆形成实验检测各组细胞的克隆形成情况，流式细胞术检测各组细胞的凋亡，MDC荧光染色检测细胞的自噬情况，Western blotting 检测自噬相关蛋白、溶酶体相关膜蛋白Lamp-2 以及组织蛋白酶Cathepsin B的表达情况。结果：顺铂耐药细胞株A2780/DDP 中Beclin1 mRNA及蛋白的表达水平均明显高于A2780 细胞株（均P<0.05），在A2780细胞中加入顺铂刺激后Beclin1 蛋白的表达水平显著升高（P<0.05）。敲减Beclin1 表达可促进顺铂诱导的A2780/DDP 细胞的凋亡（P<0.05）、抑制细胞自噬的发生（P<0.05）、减少细胞克隆形成（P<0.05）和增加细胞对顺铂的敏感性（P<0.05）；Western blotting结果显示，敲减Beclin1 可上调A2780/DDP 细胞中cleaved-caspase 3 和Cathepsin B的蛋白水平，下调Atg3、Atg7、LC3Ⅱ/Ⅰ、Lamp-2的表达水平（均P<0.05）。结论：敲减Beclin1 表达可提高A2780/DDP细胞对顺铂的敏感性，其机制可能与调节自噬相关蛋白表达抑制细胞保护性自噬和影响溶酶体功能，从而促进顺铂诱导耐药细胞凋亡有关。     

siRNA干扰MT1H基因对A549/DDP细胞耐药性的逆转

目的：探讨siRNA干扰金属硫蛋白1H（metallothionein 1H,MT1H）基因逆转A549/DDP细胞耐药的可行性。方法：采用RT-PCR方法检测MT1H基因在A549和其顺铂耐药株A549/DDP细胞中的表达;将针对MT1H的siRNA导入A549/DDP细胞;用RT-PCR和斑点印迹方法分析MT1H基因表达情况;MTT法观察细胞顺铂耐药性;TUNEL、流式细胞术检测顺铂诱导细胞凋亡率;免疫细胞化学分析凋亡相关蛋白Bcl-2、Bax的表达。结果：MT1H在A549/DDP细胞中高表达但不在A549细胞中表达;A549/DDP细胞转染48 h后,与对照组比较,MT1HsiRNA转染组MT1H mRNA和蛋白表达均明显下调,细胞对DDP的药物敏感性明显提高,DDP诱导凋亡率明显增加,Bcl-2表达明显下降,Bax表达无变化。结论：MT1H基因沉默能降低Bcl-2表达,增强顺铂对A549/DDP细胞凋亡诱导作用,有效逆转A549/DDP细胞耐药。     

BCSC-1基因异位表达对人小细胞肺癌细胞增殖的抑制效应

背景与目的:人乳腺癌候选抑制蛋白1(breast cancer suppressor candidate 1,BCSC-1)基因已被证实是一种新型抑癌基因,在多种肿瘤细胞均存在表达缺失的现象。该研究通过将BCSC-1基因转染至人小细胞肺癌细胞株NCI-H446,探讨BCSC-1基因异位表达对NCI-H446细胞增殖的抑制效应。方法:用PCR扩增BCSC-1 cDNA,构建真核重组表达载体pcDNA3.1/v5-HisB-BCSC-1。通过脂质体把pcDNA3.1/v5-HisB-BCSC-1和空质粒pcDNA3.1/v5-HisB转染入野生型NCI-H446细胞。以转染空质粒pcDNA3.1/v5-HisB的NCI-H446细胞为对照组,野生型NCI-H446细胞为空白对照组。采用流式细胞仪检测细胞周期;MTT法检测细胞增殖;免疫组化确认BCSC-1基因和CD44分子在NCI-H446细胞中的表达。结果:成功构建了真核重组表达载体pcDNA3.1/v5-HisB-BCSC-1,制备了BCSC-1基因异位高表达的NCI-H446稳定细胞株。细胞周期分析显示,异位表达BCSC-1的NCI-H446细胞大部分阻滞在G0/G1期,明显高于对照组和空白组(P<0.01)。MTT法检测显示,异位表达BCSC-1的NCI-H446细胞与对照组、空白组相比,生长速度明显减慢(P<0.05)。免疫组化显示异位表达BCSC-1的NCI-H446细胞CD44表达增高。结论:BCSC-1基因的异位表达对NCI-H446细胞的恶性增殖行为有明显的抑制作用,这种抑制作用可能与细胞周期阻滞和黏附分子CD44表达增高有关。     

miR-1293促进非小细胞肺癌耐药分子机制探讨

目的microRNA与非小细胞肺癌耐药性密切相关,但miR-1293在非小细胞肺癌中的作用机制仍不清楚。本研究探讨miR-1293调控Akt信号通路促进非小细胞肺癌耐药的分子机制。方法培养人肺癌A549与耐药细胞株A549/DDP,RT-qPCR检测miR-1293和抑癌基因RUNX3 mRNA表达,蛋白质印迹检测RUNX3蛋白表达。采用脂质体法将miR-1293抑制物(miR-1293inhibitor)和阴性对照RNA(miR-NC)转染A549/DDP。MTT检测细胞IC50;流式细胞仪检测细胞凋亡;蛋白质印迹检测凋亡相关蛋白Bcl-2和RUNX3;荧光素酶报告基因检测miR-1293能否调控RUNX3;qPCR检测miR-1293和RUNX3的mRNA表达;蛋白质印迹检测MDR1/ABCB1、ABCC1、RUNX3、Akt和p-Akt蛋白表达。结果A549/DDP细胞对DDP的耐药指数为7.23。miR-1293在耐药细胞株A549/DDP的表达(3.89±0.08,W=0.946,P=0.698)与A549组(1.01±0.05,W=0.938,P=0.622)相比升高,差异有统计学意义,t=74.78,P<0.001;而RUNX3 mRNA和RUNX3蛋白在耐药细胞株A549/DDP的表达与A549组相比降低,tmRNA=7.74,t蛋白=20.04,均P<0.001。与miR-NC比较,miR-1293 inhibitor组IC50降低,t=94.49,P<0.001。miR-1293 inhibitor组MDR1/ABCB1、ABCC1蛋白表达低于miR-NC,tMDR1/ABCB1=10.61,tABCC1=39.57,均P<0.001。与miR-NC比较,miR-1293 inhibitor组细胞凋亡增加,t=7.15,P<0.001。促凋亡蛋白RUNX3在miR-1293 inhibitor组表达更高,t=33.63,P<0.001;凋亡抑制蛋白Bcl-2表达更低,t=11.62,P<0.001。荧光素酶报告实验显示,在野生型RUNX33′UTR中,miR-1293 inhibitor的荧光素酶活性(6.24±0.04,W=0.911,P=0.399)高于miR-NC(3.21±0.07,W=0.879,P=0.221),t=92.06,P<0.001;提示RUNX3是miR-1293靶基因。miR-1293 inhibitor组RUNX3蛋白表达高于miR-NC,t=17.75,P<0.001;Akt1、p-Akt低于miR-NC组,tAkt1=67.55,tp-Akt=16.98,均P<0.001。结论在耐药细胞株A549/DDP中miR-1293高表达,抑制细胞凋亡,促进细胞耐药,其机制可能与靶向下调RUNX3,促进Akt信号通路激活有关。     

顺铂耐药乳腺癌细胞株的建立及FANCF 基因在耐药中的作用

[摘要] 目的：探讨三阴性乳腺癌顺铂（cisplatin, DDP)耐药细胞和敏感细胞中FA/BRCA通路关键基因FANCF的表达和功能,以及与DDP耐药的相关性。方法：DDP浓度递增法诱导建立乳腺癌细胞MDA-MB-231的DDP耐药细胞株MDA-MB-231/DDP;通过RNAi技术敲减MDA-MB-231 敏感细胞和DDP耐药细胞中FANCF,并在mRNA和蛋白水平进行敲减效果验证。CCK-8 法检测DDP耐药细胞株增殖活性,Western blotting 法检测该细胞中FANCF蛋白表达,流式细胞仪检测MDA-MB-231细胞周期和凋亡情况,实时定量PCR（RT-qPCR）法检测FANCF mRNA的表达。结果：MDA-MB-231细胞DDP诱导3个月建立的MDA-MB-231/DDP细胞株耐药指数为13.5,其G0/G1 期细胞增多、S期和G2/M期细胞减少。MDA-MB-231/DDP细胞中FANCF mRNA和蛋白表达水平显著升高（均P<0.01）。FANCF敲低后MDA-MB-231/DDP细胞凋亡增加,细胞对DDP的药物敏感性显著升高（均P<0.01）。结论：FANCF基因通过抗凋亡作用导致MDA-MB-231细胞对DDP的耐药性,FANCF是乳腺癌治疗的一个潜在靶点。     

RNA干扰下调拓扑异构酶Ⅰ在小细胞肺癌细胞株的表达及其与拓扑替康敏感性的关系

目的 检测拓扑异构酶Ⅰ(TopI)在小细胞肺癌(SCLC)细胞株中的表达,并探讨其表达水平对拓扑替康(TPT)敏感性的影响.方法 采用Western blot检测TopⅠ在SCLC细胞株H446蛋白水平的表达;应用人工合成并荧光标记的小干扰RNA(siRNA),通过脂质体瞬时转染H446细胞.流式细胞仪检测转染效率;应用定量逆转录聚合酶链反应(RT-PCR)和Western blot检测各干扰序列对TopI在mRNA及蛋白水平的干扰效果,筛选最佳干扰序列;对转染后的细胞株进行药物敏感性实验,观察TopI的表达对TPT敏感性的影响.结果 TOPI基因在H446中有较高表达;siRNA干扰效果满意,转染效率可达86.7%左右;siRNA oligo TopI-892、siRNA oligo TopI-1152、siRNA oHgo TopI-2084和siRNA oligo TOpI-2397等4个干扰序列均抑制H446细胞%pI mRNA的表达,抑制率分别为(95.7 ±1.6)%、(90.8 ±1.6)%、(96.1±2.7)%和(96.3±1.8)%.序列siRNA oligo TopI-2084和siRNA oligo TopI-239r7干扰后,H446细胞TopI蛋白表达明显降低.药物敏感性实验结果显示,相同浓度的TPT对实验组H446细胞的抑制率明显低于转染前及阴性对照的H446细胞(P<0.01).结论 脂质体介导的人工合成siRNA瞬时转染可有效抑制H446细胞的TOPI表达,明显降低细胞对TPT的敏感性.     

RNA干扰抑制ERCC1/ERCC2表达对非小细胞肺癌化疗敏感性影响观察

目的探讨利用RNA干扰技术沉默非小细胞肺癌A549和A549/顺铂(DDP)细胞株ERCC1和ERCC2的表达对DDP化疗敏感性的影响。方法设计并合成靶向基因siRNA-ERCC1和siRNA-ERCC2,并构建载体,通过前期转染A549/DDP细胞,筛选出沉默ERCC1和ERCC2基因表达最优的小分子RNA片段及最佳的作用时间,再将筛选出来的最优片段通过脂质体LipofectamineTM2000转染入A549细胞,观察其转染效率,使用RT-PCR及细胞免疫组化分别检测转染后基因及蛋白的表达变化;利用MTT法检测转染后细胞IC50,观察其对DDP耐药性的变化;再根据实验结果,从A549和A549/DDP细胞株中选定一株能够被逆转耐药的细胞接种于12只裸鼠皮下,观察裸鼠成瘤时间,并加用DDP局部化疗4次,记录肿瘤体积变化情况。结果通过siRNA-ERCC1和siRNA-ERCC2转染A549细胞株后,其ERCC1mRNA相对表达量为104.097 8±8.551 3,未转染组为128.465 6±12.360 4;ERCC2 mRNA相对表达量为90.415 7±7.882 1,未转染siRNA-ERCC2组为123.310 0±10.999 2。转染组的mRNA相对表达量均小于未转染组,P<0.05。转染A549细胞后,转染组ERCC1和ERCC2蛋白表达量分别为1.982 1±0.518 7和2.087 1±0.655 2,明显低于未转染组ERCC1(4.576 4±1.271 1)和ERCC2(5.680 0±1.416 7),P<0.01;MTT结果提示,siRNA-ERCC1和siRNA-ERCC2转染细胞A549后的IC50分别为(20.322 7±0.452 1)和(20.394 7±0.479 4)μg/mL,与未转染组(21.062 4±0.766 4)μg/mL差异无统计学意义,P>0.05;转染A549细胞后未发现逆转DDP耐药的效果,故筛选出A549/DDP细胞株。siRNA-ERCC1转染组的肿瘤体积为(144.63±7.67)mm3,siRNA-ERCC2转染组为(130.65±8.45)mm3,均明显小于未转染组的(252.35±12.36)mm3,P<0.01。结论高效的特异性siRNA能抑制非小细胞肺癌细胞中ERCC1和ERCC2基因和蛋白的表达,并能使A549/DDP细胞株对DDP的耐药性降低,但不能改变A549细胞株对DDP的耐药性。     

冰片逆转小细胞肺癌顺铂耐药的研究

摘 要：［目的］ 评价冰片对小细胞肺癌（SCLC）顺铂（DDP）耐药的逆转作用及其对P糖蛋白（P-glycoprotein，P-gp）和小凹蛋白-1 (Caveolin-1)表达的影响。［方法］ 利用前期建立的SCLC DDP耐药细胞株LTEP-P/DDP，四甲基偶氮唑盐微量酶反应比色法（MTT）检测冰片和/或DDP的抗肿瘤活性，流式细胞仪检测细胞周期和凋亡率，Western blotting检测P-gp和Caveolin-1的表达。［结果］冰片联合DDP对LTEP-P/DDP-0.75 细胞的增殖抑制作用较单用DDP显著性增加（P<0.05），耐药逆转指数为2.246。LTEP-P/DDP-0.75细胞经冰片联合DDP处理细胞后，较单用DDP处理组的G0/G1期和S期降低，G2/M期升高，差异有显著性（P<0.05）。冰片和DDP联合处理后，LTEP-P/DDP-0.75细胞凋亡显著性上升（P<0.05），LTEP-P/DDP-0.75细胞Caveolin-1相比LTEP-P细胞显著性上升（P<0.05），冰片或DDP单独用药组Caveolin-1显著性变化，冰片联合联合DDP组相比对照组能显著性降低LTEP-P/DDP-0.75细胞Caveolin-1表达（P<0.05），而对P-gp无显著性抑制作用。［结论］ 冰片能在一定程度上逆转LTEP-P/DDP-0.75对DDP的耐药，其作用可能与抑制Caveolin-1表达有关。     

SOX4对非小细胞肺癌细胞A549的顺铂耐药作用的影响

背景与目的 肺癌是最严重的恶性肿瘤之一,其中非小细胞肺癌发病率在肺癌中居首位.部分患者对顺铂耐受是导致化疗失败的主要原因.SOX4是转录调节因子,在多种肿瘤的发生发展过程中发挥着重要的作用,通过调控β-catenin的表达对Wnt信号通路进行调控.本研究旨在探讨SOX4对非小细胞肺癌细胞A549的顺铂耐药作用的影响.方法 体外诱导法建立顺铂耐药的肺癌细胞株A549/DDP,CCK8法检测A549及A549/DDP细胞对顺铂的耐药性.绘制A549与A549/DDP细胞生长曲线.Western blot检测耐药细胞株中SOX4的蛋白表达水平.CCK8法检测敲减SOX4后耐药细胞株A549/DDP对顺铂耐药性.实时定量PCR和免疫印迹法检测敲减SOX4后A549/DPP细胞中β-catenin和Survivin蛋白的表达.结果 成功构建非小细胞肺癌顺铂耐药细胞株A549/DDP,其耐药性是亲本细胞的13.7倍,差异有统计学意义(P<0.001).生长曲线显示耐药细胞株与亲本细胞增殖速度没有统计学差异(P<0.05);与A549细胞相比,耐药细胞A549/DDP细胞中SOX4的表达显著增高(P<0.001).敲减SOX4后,A549/DDP细胞的耐药性显著降低;敲减SOX4后,A549/DDP细胞中β-catenin和Survivin的mRNA和蛋白表达水平显著降低.结论 SOX4的表达水平影响非小细胞肺癌细胞A549对顺铂的耐药性.     

lncRNA PVT1 在结直肠癌组织和细胞中的表达及其对顺铂化疗敏感性的影响及机制

[摘要] 目的: 探讨长链非编码RNA（lncRNA）浆细胞瘤转化迁移基因1（plasmacytoma variant translocation 1, PVT1）对结直肠癌（colorectal cancer,CRC）顺铂（cisplatin,DDP）化疗敏感性的调控作用及其机制。方法: 收集2006 年4 月至2011 年3 月新乡医学院第一附属医院112 例接受手术切除、经病理确诊CRC患者的癌及癌旁组织标本,从中分别选择各30 例顺铂敏感、顺铂耐药癌及癌旁组织;人CRC细胞系HT29、SW480、HCT116、RKO和LoVo 与正常结肠上皮细胞株NCM460,构建顺铂耐药LoVo/DDP及RKO/DDP细胞。用脂质体2000 分别将siPVT1 和siNC、LV-PVT1 和LV-NC 转染或感染LoVo 和RKO细胞或者LoVo/DDP及RKO/DDP细胞。qPCR检测CRC组织及细胞中lncRNA PVT1 的表达水平。CCK-8 法、流式细胞术、WB实验分别检测敲降PVT1或过表达PVT1 对CRC细胞的增殖、凋亡及凋亡相关蛋白的表达影响。构建无胸腺裸鼠CRC皮下移植动物模型,观察对移植瘤体细胞生长及顺铂耐药的影响。结果: PVT1 mRNA在CRC组织及细胞中lncRNA PVT1 高表达,其表达水平与顺铂耐药呈正相关。敲除PVT1 后,显著降低顺铂耐药的CRC细胞的增殖能力并促进其凋亡（P<0.05 或P<0.01）、降低耐药相关分子MDR1 和MRP1 及抗凋亡相关分子Bcl-2 的表达而增加了促凋亡相关分子Bax 和活化的caspase-3 的表达。过表达PVT1 后,则促进细胞增殖并减少其凋亡（P<0.05 或P<0.01）。体内实验表明,在CRC细胞中过表达PVT1 促进顺铂耐药的形成（P<0.05）。结论: 敲降lncRNA PVT1 表达可显著抑制CRC耐药细胞株的增殖并促进其凋亡,过表达PVT1 可明显促进CRC细胞和动物移植瘤体的生长。PVT1 通过抑制MDR1 和MRP1 的表达,调控内源性凋亡通路,进而增强CRC细胞对顺铂的敏感性。     

Methylation Profiles of BRCA1, RASSF1A and GSTP1 in Vietnamese Women with Breast Cancer

Objective: This study investigated the DNA promoter methylation profiles of BRCA1, RASSF1A and GSTP1 genes,both individually and in an integrative manner in order to clarify their correlation with clinicopathological parameters ofbreast cancer from Vietnamese patients, and establish new potential integrative methylation biomarkers for breast cancerdetection. Material and methods: The methylation frequencies of BRCA1, RASSF1A and GSTP1 were analyzed bymethylation specific polymerase chain reaction (MSP) in 70 specimens of breast carcinomas and 79 pairs of tumor andmatched adjacent normal tissues from breast cancer patients. Results: All the three analyzed genes showed a concordanceconcerning their promoter methylation in tumor and adjacent normal tissue. The methylation of BRCA1, RASSF1Aand GSTP1 was found in 58.23 %, 74.68 % and 59.49 % of tumor tissues and 51.90 %, 63.29 % and 35.44 % ofcorresponding adjacent tissues, respectively. When each gene was assessed individually, only the methylation ofGSTP1 was significantly associated with tumor tissues (p=0.003). However, the methylation frequency of at least one ofthe three genes and the methylation frequency of all the three genes both showed significant association with tumor(p=0.008 and p=0.04, respectively). The methylation of BRCA1 was found to be significantly associated with tumorgrade (p=0.01). Conclusion: This study emphasized that the panel of the three genes BRCA1, RASSF1A and GSTP1can be further developed as potential biomarkers in diagnosis and classification of breast cancer in Vietnamese women.     

Multiplex PCR with Confronting Two-pair Primers for CYP1A1 Ile462Val,GSTM1, GSTT1, and NQO1 C609T

Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) is an effective genotyping method ‍for single nucleotide polymorphisms (SNPs) in aspects of reducing time and costs for analysis. So far we have ‍established PCR-CTPP conditions for tens of SNPs, including a triplex genotyping (Kawase et al., 2003). In the ‍present study we report a quadruplex PCR-CTPP to genotype simultaneously four functional polymorphisms of ‍carcinogen-metabolizing enzymes, CYP1A1 Ile462Val, GSTM1 null, GSTT1 null and NQO1 C609T, which were ‍reported that they have significant associations with smoking-related cancers. We applied this method for 475 health ‍check-up examinees to demonstrate the performance. Among the subjects, the genotype frequency of CYP1A1 ‍Ile462Val was 56.8% for Ile/Ile, 38.1% for Ile/Val and 5.1% for Val/Val. The null type frequencies of GSTM1 and ‍GSTT1 were 52.8% and 49.9%, respectively. And the genotype frequency of NQO1 C609T was 41.9% for C/C, ‍41.3% for C/T and 16.8% for T/T. Their distributions were similar to those reported for Japanese by other studies. ‍To the best of our awareness, this is the first paper that reports the success in quadruplex PCR-CTPP. The applied ‍polymorphisms are useful ones, which would be adopted not only for research purposes, but also for risk assessment ‍of individuals exposed to carcinogenic substances. This convenient genotyping would be applied for cancer prevention ‍especially in Asian Pacific regions, where expensive genotyping methods are hardly available.     

Expression of seven-in-absentia homologue 1 and hypoxia-inducible factor 1 alpha: Novel prognostic factors of nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is an EBV-associated cancer. We analysed Siah1 expression as well as LMP1 and HIF1α expression by immuno-histochemical staining in 74 NPC biopsy specimens and found that the expression of Siah1 was significantly correlated with advanced tumour status and stage. Moreover, Siah1-positive and HIF1α-positive cases had significantly worse prognoses. The expression score for LMP1 was remarkably correlated with that of Siah1, whereas there was little correlation between LMP1 expression and the other markers evaluated. This is the first study to evaluate the pattern and clinical significance of Siah1 and HIF1α expression in NPC, and such an evaluation is valuable for identifying those patients at a high risk for a poor prognosis.     

黏蛋白1多肽通过small-MUC1蛋白抑制肿瘤细胞生长

目的：探讨黏蛋白1（mucin 1,MUC1）串联重复序列多肽(简称黏蛋白1多肽,MUC1多肽)对肿瘤细胞生长抑制的作用机制。方法：MUC1多肽与多种肿瘤细胞Jurkat、Raji、U937、MCF7、SMMC7721及活化的T细胞、小鼠巨噬细胞RAW264.7共同培养,观察MUC1多肽对上述细胞生长的影响;建立BABL/c小鼠Jurkat细胞皮下移植瘤动物模型,应用MUC1多肽进行治疗;采用GST免疫沉降实验鉴定与MUC1多肽结合的肿瘤细胞表面蛋白。结果：MUC1多肽对Jurkat、Raji、U937、MCF7和SMMC7721细胞的生长均有抑制作用,对活化的T细胞和小鼠RAW264.7细胞生长无明显抑制作用。MUC1多肽对BABL/c小鼠皮下Jurkat细胞移植瘤的生长均有明显抑制作用（P<0.05）。GST免疫沉降实验显示,Jurkat 和MCF7细胞裂解上清中与MUC1多肽结合的蛋白可与两种抗MUC1串联重复序列抗体（GP1.4和HMPV）及抗胞内段抗体（Ab5）发生反应,相对分子质量大约115 000,提示可能是MUC1新的同种型,命名为small MUC1（sMUC1）。结论：MUC1多肽可通过与肿瘤细胞表面small MUC1蛋白的相互作用向细胞传导生长抑制信号     

Upregulated long non-coding RNA AFAP1-AS1 expression is associated with progression and poor prognosis of nasopharyngeal carcinoma

Altered expression of long noncoding RNAs (lncRNAs) associated with human carcinogenesis. We performed a cDNA microarray analysis of lncRNA expression in 12 cases of nasopharyngeal carcinoma (NPC) and 4 non-tumor nasopharyngeal epitheliums. One lncRNA, actin filament associated protein 1 antisense RNA1 (AFAP1-AS1), was identified and selected for further study. AFAP1-AS1 expression was upregulated in NPC and associated with NPC metastasis and poor prognosis. In vitro experiments demonstrated that AFAP1-AS1 knockdown significantly inhibited the NPC cell migration and invasive capability. AFAP1-AS1 knockdown also increased AFAP1 protein expression. Proteomic and bioinformatics analyses suggested that AFAP1-AS1 affected the expression of several small GTPase family members and molecules in the actin cytokeratin signaling pathway. AFAP1-AS1 promoted cancer cell metastasis via regulation of actin filament integrity. AFAP1-AS1 might be a potential novel marker that can predict cancer patient prognosis and as a potential therapeutic target for NPC.     

The lncRNA FEZF1-AS1 Promotes the Progression of Colorectal Cancer Through Regulating OTX1 and Targeting miR-30a-5p

Long noncoding RNAs (lncRNAs) participate in and regulate the biological process of colorectal cancer (CRC) progression. Our previous research identified differentially expressed lncRNAs in 10 CRC tissues and 10 matched nontumor tissues by next-generation sequencing (NGS). In this study, we identified an lncRNA, FEZF1 antisense RNA 1 (FEZF1-AS1), and further explored its function and mechanism in CRC. We verified that FEZF1-AS1 is highly expressed in CRC tissues and cell lines. Through functional experiments, we found that reduced levels of FEZF1-AS1 significantly suppressed CRC cell migration, invasion, and proliferation and inhibited tumor growth in vivo. Mechanistically, we discovered that reduced levels of the lncRNA FEZF1- AS1 inhibited the activation of epithelial–mesenchymal transition (EMT); the overexpression of orthodenticle homeobox 1 (OTX1) partially rescued the FEZF1-AS1-induced inhibition of protein expression. It indicated that FEZF1-AS1 may play a role in the occurrence and development of CRC by regulating the FEZF1-AS1/ OTX1/EMT pathway. Furthermore, it was reported that FEZF1-AS1 is located in both the nucleus and cytoplasm of HCT116 cells. Dual-luciferase reporter assays verified that FEZF1-AS1 directly binds miR-30a-5p and negatively regulated each other. Further, we showed that 5 -nucleotidase ecto (NT5E) is a direct target of miR-30a-5p, and the inhibition of miR-30a-5p expression partially rescued the inhibitory effect of FEZF1-AS1 on NT5E. Our results indicated that the mechanism by which FEZF1-AS1 positively regulates the expression of NT5E is through sponging miR-30a-5p. Our study demonstrated that lncRNA FEZF1-AS1 is involved in the development of CRC and may serve as a diagnostic and therapeutic target for CRC patients.     

Methylation Profile of BRCA1, RASSF1A and ER in Vietnamese Women with Ovarian Cancer

DNA methylation is considered a promising biomarkers for diagnosis of cancer in general and of ovariancancer in particular. In our study, we validated the accuracy of methylation specific polymerase chain reaction(MSP) to analyze the methylation pattern of BRCA1, RASSF1A and ER in 59 and 10 Vietnamese patients withepithelial ovarian cancer (EOC) and benign ovarian tumors, respectively. We found methylation of BRCA1,RASSF1A and ER in 11/59 (18.6%), 40/59 (67.8%) and 15/59 (25.4%) of EOC cases, while methylation of BRCA1was only detected in 2/10 (20%) benign ovarian patients. Forty five out of the 59 EOCs (78%) demonstratedmethylation at one or more genes. The methylation frequency of RASSF1A was significantly associated with EOC(p<0.0005). No significant association was observed between methylation status of these genes and the clinicaland pathological parameters of tumors collected from Vietnamese women suffering from ovarian cancer.     

免疫检查点PD-1／PD-L1通路在小细胞肺癌中的作用

小细胞肺癌（SCLC）主要的特点为生长迅速且早期易发生广泛转移。尽管SCLC对于化疗和放疗敏感,但几乎所有的患者均在治疗后发生复发转移,预后差。免疫检测点抑制剂,尤其程序化细胞死亡受体-1（PD-1）／程序化细胞死亡配体-1（PD-L1）拮抗剂在SCLC的临床前和临床研究中均获得了良好效果,并且能够延长患者生存。免疫检测点疗法作为一种新兴的方法在未来可能会改变SCLC治疗模式。此外,有限的数据显示出PD-L1表达可能成为筛选获益人群一个有效的生物标记物。本文总结PD-L1作为标记物的发展历程,并同时阐述PD-1／PD-L1抑制剂在SCLC治疗中的进展。     

Glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) Polymorphisms and Lung Cancer Risk among a Select Group of Iranian People

Objective(s): Lung cancer, caused primarily by smoking, is one of the leading determinants of mortality throughoutthe world. Here we investigated the effects of polymorphisms in two enzymes, i.e., GSTT1 and GSTM1, related tothe antioxidant defense line against carcinogens associated with lung cancer among a select group of Iranian people.Materials and Methods: One hundred and twenty lung cancer patients from two referral centers in Tehran, Iran, wererecruited for comparison with 120 healthy controls. Genomic DNA was extracted from the FFPE tumor tissues ofthe select cases and peripheral blood buffy coats of healthy controls. The polymorphisms of GSTT1 and GSTM1 wereinvestigated by multiplex polymerase chain reaction. Results: With the 240 samples studied, no specific relationshipwith lung cancer was discerned for the GSTM1 (P=0.35; OR=1/33; 95% CI=0.79-2.25) polymorphism, but the GSTT1(P=0.005; OR=2.4; CI=1.32-4.35) gene polymorphism revealed a notable association on logistic regression, takinginto account age and sex factors. Furthermore, the GSTT1 genotype distribution in patients with LSCC was differentfrom that of healthy cases (P=0.006; OR=3.11; CI=1.38-7.04). The risk of developing lung cancer with the T0M1genotype was 3.46 times higher than with T1M1 genotype (P=0.002; OR=3.46; CI=1.61-7.46). Moreover, the risk ofdeveloping LSCC cancer in people with T0M1 genotypes was significantly elevated (P=0.004; OR=4.5; CI=1.62-12.52).Conclusion: Unlike GSTM1, the GSTT1 genotype distribution is associated with the incidence of lung cancer in Iranianpeople. Different types of lung cancer appear to show various correlations with GST polymorphisms in this regard.