阿帕替尼对人肝癌细胞增殖及凋亡的作用机制研究

目的:探讨阿帕替尼抑制肝癌细胞增殖促进凋亡的作用机制。方法:选取肝癌细胞系SNU739、HepG2,以CCK-8细胞增殖实验、平板克隆实验测定阿帕替尼对肝癌细胞增殖及克隆形成能力的影响;流式细胞术检测阿帕替尼对肝癌细胞凋亡的影响;蛋白免疫印迹法检测阿帕替尼影响肝癌细胞凋亡相关蛋白Bax、Bcl-2及Caspase3的表达情况。结果:与对照组相比,阿帕替尼可显著抑制肝癌细胞增殖(P0.05)。平板克隆实验提示与对照组相比,10μM和20μM阿帕替尼组肝癌细胞克隆数明显减少(P0.05)。流式细胞术结果提示10μM和20μM阿帕替尼处理组细胞凋亡率明显增加(P0.05)。蛋白免疫印迹法结果显示经阿帕替尼处理的肝癌细胞,促凋亡蛋白Bax及Caspase3的活性片段Cleaved-caspase3表达水平显著上调,抗凋亡蛋白Bcl-2显著下调(P0.01)。结论:阿帕替尼通过调节肝癌细胞凋亡相关蛋白从而抑制肝癌细胞增殖、促进其凋亡。     

长非编码RNA SNHG18对人胃癌细胞BGC823增殖和凋亡的影响

目的:探究长非编码RNA SNHG18对胃癌细胞增殖和凋亡的影响。方法:采用实时定量PCR(qRT-PCR)技术检测人胃癌组织及癌旁组织和胃癌细胞系中lncRNA SNHG18的表达;采用MTT和克隆形成试验观察转染SNHG18过表达质粒后胃癌细胞BGC823增殖活力的变化;通过流式细胞术检测lncRNA SNHG18对胃癌细胞BGC823凋亡的影响。结果:相较于癌旁组织和胃正常粘膜上皮细胞系GSE-1,胃癌组织及胃癌细胞系中SNHG18的表达水平显著降低(P0.05);胃癌细胞过表达SNHG18增殖活力以及克隆形成的能力均显著降低(P0.05),而细胞凋亡率明显升高(P0.05)。结论:胃癌组织中长非编码RNA SNHG18呈低表达,可促进胃癌细胞增殖并抑制其凋亡,可能在胃癌发生发展过程中发挥重要作用。     

hMMS2基因对结肠癌细胞耐药逆转的影响

为探讨hMMS2(Human methyl methanesulfonate sensitive mutant 2)基因对人结肠癌细胞耐药逆转的影响, 文章以人高分化耐奥沙利铂结肠癌细胞(THC8307/L-OHP)为实验材料, 采用脂质体-质粒转染技术构建了带有干扰目的基因hMMS2的miRNA片段并携带绿色荧光蛋白标记重组质粒(pcDNA6.2-GW/EmGFP-miR-MMS2)的细胞系, 通过实时荧光定量PCR(qRT-PCR)和免疫荧光技术(Immunostaining technique)检测该细胞系的干扰效率。选择hMMS2低表达具有统计学意义的上述细胞系作为实验组细胞, 同时将未曾作过处理的THC8307/ L-OHP细胞作为空白对照组, 转染绿色荧光蛋白空质粒(pcDNA6.2-GW/EmGFP-miR)的THC8307/L-OHP细胞作为阴性对照组, 以噻唑蓝比色分析实验(MTT colorimetric analysis assay)、克隆形成实验(Colony formation assay)对3组细胞的存活率和克隆形成率进行检测, 结果显示：实验组细胞的奥沙利铂半数抑制浓度(Half inhibition concentration, IC50)、耐药指数(Resistance index, RI)及克隆形成率(Colony-forming efficiency, CFE)均比对照组细胞明显降低(P＜0.05), 而相对逆转率(Relative reverse efficiency, RRE)增高(P＜0.05), 提示实验组细胞增殖能力减弱; 以罗丹明123实验(Rhodamine 123 assay)结合倒置荧光显微镜、流式细胞仪检测技术等观测细胞的凋亡变化, 结果显示, 实验组细胞的凋亡率较对照组细胞显著增高(P＜0.05); 两对照(空白、阴性)组间并无细胞增殖或凋亡的显著性差异。研究结果提示：下调hMMS2基因表达可逆转人高分化耐奥沙利铂结肠癌细胞对L-OHP的耐药性并促进结肠癌细胞的凋亡。     

利用Tet-On系统敲低PRDM5对BEAS-2B细胞增殖的影响

[目的]探讨利用dox诱导的Tet-On调控系统敲低PRDM5基因对人正常支气管上皮细胞BEAS-2B增殖的影响。[方法]构建敲低PRDM5基因的p LKO-Tet-On-PRDM5-shRNA慢病毒质粒,包装生产慢病毒并转染BEAS-2B细胞,以相同条件制备转染p LKO-Tet-On-control-shRNA的BEAS-2B细胞作为对照。Western Blot法检测PRDM5蛋白的表达水平; MTT实验观察细胞增殖情况;平板克隆形成实验检测细胞增殖能力;流式细胞术检测细胞凋亡水平。[结果]在合适剂量的dox诱导下(0. 5μg/mL),敲低组细胞中PRDM5蛋白表达明显低于对照组(P 0. 05);各时间点OD值高于对照组(P 0. 05);克隆形成数大于对照组(P 0. 01);凋亡率低于对照组(P 0. 01)。[结论]成功构建出Tet-On系统调控敲低PRDM5的BEAS-2B细胞系。PRDM5基因敲低可促进BEAS-2B细胞增殖并且抑制其凋亡。     

蒿本内酯通过lncRNA NKILA对干扰素α诱导的人脑血管外膜成纤维细胞增殖及凋亡的影响

该文旨在探讨蒿本内酯对干扰素α(IFN-α)诱导的人脑血管外膜成纤维细胞(HBVAF)增殖及凋亡的影响及其可能作用机制。HBVAF细胞经IFN-α诱导后建立细胞损伤模型,将不同剂量的蒿本内酯作用于IFN-α诱导的HBVAF细胞后,采用MTT法、平板克隆形成实验、流式细胞术分别检测细胞增殖、克隆形成及凋亡情况, qRT-PCR法检测lncRNA NKILA的表达量;为探究蒿本内酯与lncRNA NKILA对IFN-α诱导的HBVAF细胞增殖及凋亡的影响,将pcDNA、pcDNA-lncRNA NKILA分别转染至HBVAF细胞后加入IFN-α处理24 h, si-NC、si-lncRNA NKILA分别转染至HBVAF细胞后加入蒿本内酯与IFN-α共处理24 h, Western blot测定凋亡相关蛋白表达量。结果显示,蒿本内酯处理后细胞存活率和lncRNA NKILA的表达量升高(P<0.05),细胞克隆形成数增多(P<0.05),细胞凋亡率和Cleaved-caspase3、Cleaved-caspase9蛋白水平降低(P<0.05),且呈剂量依赖性;转染pcDN...     

ADAR1 shRNA对人胶质瘤U87细胞增殖和凋亡的影响

目的:研究ADAR1 shRNA对人胶质瘤细胞U87细胞增殖和凋亡的影响。方法:通过构建ADAR1-shRNA的干扰质粒,经脂质体法转染胶质瘤U87细胞系,通过荧光倒置显微镜观察转染效率,选择转染效率最高的细胞系。取转染48h细胞,采用RT-PCR和Western-blot分别检测ADAR1 mRNA及蛋白的表达,流式细胞仪检测其细胞凋亡率,MTT法检测细胞增殖情况。结果:①经ADAR1-shRNA转染48h后的转染效率最高,此时U87细胞系中ADAR1 mRNA及蛋白的表达均被显著抑制,较阴性对照组及空白组均明显降低(P0.05)。②在转染ADAR1-shRNA后,细胞凋亡率为(28.14%±3.76%),明显高于阴性对照组(3.20%±1.57%)和空白组(2.80%±1.49%),细胞增殖率较阴性对照组及空白组明显下降(P0.05)。结论:通过shRNA抑制ADAR1的表达能明显促进人胶质瘤细胞U87细胞的凋亡和抑制其增殖,ADAR1基因可能成为治疗治疗胶质瘤的新靶点。     

IL27RA对EBV感染的鼻咽癌细胞TRAF1/2表达的调控及其生物学效应

EB病毒(Epstein-Barr virus,EBV)感染是鼻咽癌(Nasopharyngeal carcinoma,NPC)最主要的危险因素,主要通过肿瘤坏死因子受体相关分子-1/2(Tumor necrosis factor receptor-associated factor-1/2,TRAF1/2)发挥促癌作用,为探究白介素27受体亚基α(Interleukin 27 receptorα,IL27RA)表达与NPC细胞EBV感染的关系及其生物学效应,本研究采用肿瘤基因组计划(The Cancer Genome Atlas,TCGA)数据库在NPC组织中筛选EBV感染相关信号分子TRAF1/2的潜在相互作用分子,培养携带EBV的NPC细胞株HK-1和C666-1及未携带EBV的NPC细胞株CNE-1及CNE-2,实时荧光定量PCR技术(RT-qPCR)及蛋白质印迹技术(Western Blot)检测IL27RA mRNA及蛋白表达,慢病毒介导细胞转染技术外源性干扰HK-1和C666-1细胞IL27RA的表达,CCK-8检测细胞增殖能力的变化情况,Transwell实验检测细胞侵袭能力的变化情况。结果显示,NPC组织中存在与TRAF1/2表达均显著正相关的分子IL27RA(相关系数R分别为0.61及0.69,P均小于0.001),同时IL27RA在NPC组织中高表达(P0.001),HK-1和C666-1中IL27RA mRNA及蛋白的表达水平显著高于CNE-1及CNE-2细胞(P0.01),干扰HK-1和C666-1细胞IL27RA表达后,TRAF1及TRAF2 mRNA及蛋白表达显著下调(P0.01),且细胞增殖及转移能力均显著减弱(P0.01)。本研究提示,干扰IL27RA表达可显著抑制携带EBV的NPC细胞TRAF1/2的表达及细胞增殖转移能力,IL27RA有望成为NPC新的分子靶标或标志物。     

丹参酮ⅡA联合CASC2对甲状腺癌细胞增殖、凋亡、迁移、侵袭的影响固有淋巴细胞在脂质代谢中的研究进展

为了研究丹参酮ⅡA联合长链非编码RNA(lnc RNA)癌易感性候选基因2(CASC2)对甲状腺癌细胞增殖、凋亡、迁移、侵袭的影响,该研究采用实时荧光定量PCR(q RT-PCR)检测CASC2在甲状腺癌组织中的表达。将甲状腺癌SW579细胞分为pc DNA3.1组(转染pc DNA3.1质粒), pc DNA3.1-CASC2组(转染pc DNA3.1-CASC2质粒), con组(用与丹参酮ⅡA等量的二甲基亚砜处理),药物-1、2、3、4组(分别用1、2、4、8μg/m L丹参酮ⅡA处理),药物-4+pc DNA3.1组(转染pc DNA3.1质粒且用8μg/m L丹参酮ⅡA处理),药物-4+pc DNA3.1-CASC2组(转染pc DNA3.1-CASC2质粒且用8μg/m L丹参酮ⅡA处理)。分别用细胞计数试剂盒(CCK-8)和平板克隆检测细胞存活与克隆形成;流式细胞术检测细胞凋亡; Transwell检测细胞迁移、侵袭;蛋白质印迹法(Western blot)检测蛋白P21、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、E-钙黏蛋白(E-cadherin)和基质金属蛋白酶-2(MMP-2)的表达。结果显示,与癌旁组织相比,甲状腺癌组织中的CASC2表达量显著降低(P0.05)。过表达CASC2明显降低SW579细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数和MMP-2蛋白表达量,显著提高细胞凋亡率、P21、Caspase-3、E-cadherin蛋白表达量(P0.05)。丹参酮ⅡA明显降低SW579细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数、MMP-2蛋白水平,显著提高细胞凋亡率、P21、Caspase-3、E-cadherin蛋白表达水平,且均呈浓度依赖性(P0.05)。丹参酮ⅡA联合CASC2明显降低SW579细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数、MMP-2蛋白表达量,显著提高细胞凋亡率、P21、Caspase-3和E-cadherin蛋白水平(P0.05)。因此,丹参酮ⅡA联合CASC2可以抑制甲状腺癌细胞的增殖、迁移和侵袭,以及诱导细胞凋亡。     

LTF基因在鼻咽癌细胞系中生物学功能的初步研究

目的:初步探讨野生型LTF基因在鼻咽癌细胞系中的生物学功能。方法:野生型LTF导入鼻咽癌细胞系,G418筛选,RT-PCR和Western-blotting分别在mRNA和蛋白质水平进行验证,得到稳定表达LTF基因的鼻咽癌细胞系。流式细胞术、平板克隆形成实验和MTT法分别检测细胞周期、细胞的克隆形成能力和细胞生长曲线。结果:成功导入LTF并稳定表达的鼻咽癌细胞系,G0-G1期细胞百分比例明显增加(72.01%vs 62.31%),G2-M期细胞百分比例减少(6.26%vs 10.81%);克隆形成能力降低(39.5%vs 59.7%),体外瘤细胞增殖能力降低(P0.05)。结论:LTF基因可阻滞细胞周期、抑制鼻咽癌细胞系的增殖能力和克隆形成率,同时为进一步的体内试验研究奠定基础。     

裂蹄木层孔菌子实体水提物诱导HepG2细胞凋亡的初步研究

研究裂蹄木层孔菌子实体水提物(WEPL)对人类克隆肝癌细胞系HepG2生长的作用。用裂蹄木层孔菌子实体水提物处理HepG2细胞后,噻唑蓝法(MTT法)可见浓度和时间依赖性抑制细胞增殖;电镜下观察凋亡小体的出现,流式细胞仪技术显示Annexin-Ⅴ染色呈阳性,都证明了HepG2细胞发生了凋亡。RT-PCR和Western Blot分析证实WEPL刺激Bax表达量上调、Bcl-2表达量下调进而诱导了细胞凋亡。结果表明WEPL诱发的克隆人类肝癌细胞系HepG2的细胞凋亡可能是通过上调Bax、下调Bcl-2活性来实现的。     

慢病毒转染对鼻咽癌细胞5-8F增殖迁移的影响

目的:探讨慢病毒转染对鼻咽癌细胞株5-8F增殖、迁移的影响,以验证慢病毒转染是否能有效的应用于鼻咽癌细胞的增值及迁移相关研究。方法:以红色荧光标记的慢病毒为转染载体,选定不同的MOI值转染鼻咽癌5-8F细胞株,扩大培养后筛选纯化,流式细胞仪检测转染效率。以最佳MOI值转染后的5-8F(RFP-5-8F)细胞为实验组,未转染的亲代5-8F为空白对照组,取对数生长期未转染的亲代5-8F和红色荧光标记的慢病毒转染的5-8F(RFP-5-8F)细胞进行MTT、划痕实验,观察细胞镜下形态,了解细胞转染前后生长曲线,细胞迁移能力的变化。结果:流式细胞仪检测5-8F细胞慢病毒转染效率大于95%,转染最佳MOI值为30,镜下荧光强度适中。实验组与对照组比较,转染前后5-8F细胞光镜形态相似,生长曲线一致,差异无统计学意义(P=0.997),划痕实验显示5-8F与RFP-5-8F细胞迁移能力一致,差异无统计学意义(P0.05)。结论:慢病毒转染后鼻咽癌细胞能真实有效的的反应原细胞的增值及迁移能力,可以很好的应用于鼻咽癌增殖及其转移机制的相关研究。     

shRNA interference for extracellular signal-regulated kinase 2 can inhibit the growth of esophageal cancer cell line Eca109

Background: Esophageal squamous cell carcinoma is one of the most common digestive tract cancers with 5-year survival rate less than 10% owing to its poor prognosis. Mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway has been mainly involved in the pathogenesis of various cancers. In present study, we investigated the role of ERK2 in human esophageal cancer cell line Eca109.

Methods: Short-hairpin RNA (shRNA) interference vector targeted ERK2 was constructed using pGeneclip U1 hairpin cloning systems, then transfected into Eca109 cell line. The transfection efficiency was observed by fluorescence microscope and cell growth after transfection with shRNA-ERK2 vector was determined by methylthiazolyl blue tetrazolium (MTT) assay. The ERK2 expression after transfection was detected by western-blotting. The cell apoptosis and cell-cycle was analyzed by flow cytometry. The role of p-ERK2 was confirmed by immunohistochemistry and soft agar colony formation assay.

Results: The growth of Eca109 transfected with shRNA-ERK2 vector was obviously inhibited compared to control group via MTT analysis. The inhibition rate after transfection with shRNA-ERK2 for 96?h was 10.45%, the expression of ERK2 was obviously reduced compared to the control analyzed by western-blot, cell apoptosis was 9.7% (compared to control, P?<?0.05), and cell-cycle was arrested at G1 phase.

Conclusions: In present study we demonstrated for the first time that transfection with shRNA-ERK2 targeted ERK2 into Eca109 cells can inhibit growth of Eca109, inducing cell apoptosis and influencing cell-cycle. Together, these results we obtained suggested that ERK2 plays an important role in cell growth of Eca109.     

miR-221对甲状腺乳头癌生物学特性的影响

目的:探讨miR-221对甲状腺乳头癌生物学特性的影响。方法:培养人甲状腺乳头癌细胞株BCPAP、K1、TPC-1和正常甲状腺细胞株Nthy-ori 3-1。将实验分为四组:A:miR-221模拟物组;B组:miR-221抑制物组;C:无关序列组;D:空白对照组。RT-q PCR的方法检测miR-221在各个细胞中的表达以及转染后各组细胞的表达;MTT实验检测转染后各组细胞的增殖;划痕实验检测转染后各组细胞的迁移能力;流式细胞仪检测转染后各组细胞的凋亡情况。结果:RT-qPCR检测miR-221在三个细胞株的表达情况显示,miR-221甲状腺乳头癌细胞株TPC-1的表达最高,因此选择TPC-1作为后续的研究;miR-221在转染后各组细胞的表达量显示,转染miR221模拟物的miR221的表达显著高于空白对照组,转染miR221抑制物的miR221的表达显著低于空白对照组(P0.001);MTT实验结果显示,转染miR-221模拟物组细胞的增殖速度最快,转染miR-221抑制物组细胞的增殖速度最慢,miR-221模拟物组和miR-221抑制物组细胞从第三天开始与空白对照组有显著差异(P0.01),无关对照组与空白对照组无显著差异(P0.05);划痕实验结果显示,转染miR-221模拟物组细胞的迁移数显著高于空白对照组,转染miR-221抑制物组细胞的迁移数显著低于空白对照组(P0.01),无关对照组与空白对照组无显著差异(P0.05);流式细胞仪结果显示,转染miR-221模拟物组细胞凋亡率显著低于空白对照组(P0.01),转染miR-221抑制组细胞凋亡率显著高于空白对照组(P0.001),转染无关对照对细胞凋亡无影响(P0.05)。结论:过表达miR-221可促进细胞增殖、迁移,抑制细胞凋亡。抑制miR-221表达可降低细胞增殖、迁移,增加细胞凋亡。     

RNA干扰NUP88基因对人乳腺癌MCF-7细胞生长及侵袭力的影响

目的： 构建重组慢病毒介导的NUP88-shRNA载体,通过RNAi技术分别观察沉默NUP88后对MCF-7增殖,粘附,侵袭和转移情况的影响,为乳腺癌的临床基因治疗寻找新的靶点。方法： 构建NUP88重组慢病毒表达载体,包装后检测滴度,以最佳复感染指数转染乳腺癌MCF-7细胞,利用RT-PCR和Western blot检测各组MCF-7细胞中mRNA和蛋白的表达效率;MTT法和流式细胞仪检测法,检测NUP88基因被干扰后对MCF-7细胞增殖和凋亡的影响;细胞侵袭实验检测NUP88基因被干扰后对MCF-7侵袭力的影响。结果 四组病毒及一组阴性对照均构建成功,滴度均为4E+8TU/ml;RT-PCR和Western blot检测,结果表明：经NUP88-shRNA转染的MCF-7细胞组NUP88 mRNA和蛋白质的表达与经阴性转染组和空白MCF-7细胞组相比,差异明显具有统计学意义（P<0.01）;测定NUP88-shRNA1组沉默效率最高,沉默率可达到86%;MTT法结果表明：实验组经NUP88-shRNA1慢病毒转染后细胞增殖程度显著减少,与空白组和对照组相比有显著性差异（P<0.05）。流式细胞仪检测三组MCF-7细胞凋亡结果表明：实验组经慢病毒转染后细胞凋亡率显著增加,与对照组和空白组相比有显著性差异（P<0.05）;细胞侵袭实验表明：在肿瘤细胞常规培养24h后,实验组与空白组和阴性对照组比较,穿膜细胞数量明显减少,具有显著性差异（P<0.05） 结论： NUP88重组慢病毒可以通过RNAi成功抑制MCF-7中NUP88基因的表达,并能显著抑制其增殖及远处的侵袭能力。     

Impact of BTG2 expression on proliferation and invasion of gastric cancer cells in vitro

BTG2 (B cell translocation gene 2) is downregulated in several human tumors and has been known as a tumor suppressor in carcinogenesis of thymus, prostate, kidney, and liver. However, little is known about the role BTG2 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of BTG2 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell lines SGC7901 and MKN45. BTG2 cDNA was insected into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 and SGC7901 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these transfectants were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay, respectively. The invasion of these clones was analyzed by using cell migration assay. MKN-BTG2 (MKN45 with stable transfection of BTG2 gene) and SGC-BTG2 (SGC7901 with stable transfection of BTG2 gene) grew slower than their control groups, respectively. The cell counts of MKN-BTG2 in the fourth, fifth, sixth and seventh days were significantly fewer than those of control groups (P < 0.05). Those of SGC-BTG2 in the fourth fifth, sixth and seventh days were significantly fewer than those of control groups too (P < 0.05). Cell cycle analysis showed that proportions of MKN-BTG2 and SGC-BTG2 cells in G0–G1 and S were different significantly with those of their control groups, respectively (P < 0.05). The apoptosis rate of MKN-BTG2 was significantly higher than those of control groups (P < 0.05). Results of colony-forming assay showed that the colon formation rates of MKN-BTG2 and SGC-BTG2 were lower than those of their control groups (P < 0.05). The results of cell migration assay showed that the cell migration rates of MKN-BTG2 and SGC-BTG2 were not significantly different with those of their control groups (P > 0.05). BTG2 can restrain the growth and proliferation of gastric cancer cells powerfully. It can reduce some malignant phenotype of these tumor cells. But it could not impact the ability of invasion of gastric cancer cells, so could not restrain the metastasis of gastric cancer. In gastric cancer, BTG2 could be thought as a tumor-inhibiting gene in some distance, so the gene could be a potential target of gene therapy.     

Silencing of long noncoding RNA SRRM2-AS exerts suppressive effects on angiogenesis in nasopharyngeal carcinoma via activating MYLK-mediated cGMP-PKG signaling pathway

Long noncoding RNAs (lncRNAs) play a crucial role in several malignances, involving nasopharyngeal carcinoma (NPC), a heterogeneous disease. This study investigated mechanism of serine/arginine repetitive matrix protein 2-alternative splicing (SRRM2-AS) in NPC cell proliferation, differentiation, and angiogenesis. Initially, differentially expressed lncRNAs were screened out via microarray analysis. Vascular endothelial growth factor (VEGF) protein positive rate and microvessel density (MVD) were determined in NPC and adjacent tissues. NPC CNE-2 cells were treated with a series of vector and small interfering RNA to explore the effect of SRRM2-AS in NPC. The target relationship between myosin light chain kinase (MYLK) and SRRM2-AS was verified. Levels of SRRM2-AS, MYLK, cGMP, PKG, VEGF, PCNA, Ki-67, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase 3 were determined after transfection. Finally, the effect of SRRM2-AS on cell proliferation, colony formation, angiogenesis, cell cycle, and apoptosis in NPC was evaluated. SRRM2-AS was highly expressed and MYLK was poorly expressed in NPC tissues. VEGF protein positive rate and MVD were elevated in NPC tissues. MYLK was confirmed to be a target gene of SRRM2-AS. Silencing of SRRM2-AS elevated levels of MYLK, cGMP, PKG, Bax, and Caspase 3, but decreased levels of VEGF, PCNA, Ki-67, and Bcl-2. Especially, silencing of SRRM2-AS suppressed cell proliferation, colony formation and angiogenesis, blocked cell cycle, and enhanced cell apoptosis in NPC. Our results suggested that silencing of SRRM2-AS protected against angiogenesis of NPC cells by upregulating MYLK and activating the cGMP-PKG signaling pathway, which provides a new target for NPC treatment.     

穿膜素TAT介导的KDR-siRNA慢病毒载体的构建及其靶向肺癌A549细胞的抗肿瘤作用

为了构建TAT与KDR-siRNA慢病毒载体,观察其对肺癌细胞株 A549的体外靶向抗肿瘤作用,利用重组技术构建TAT-KDR siRNA慢病毒载体并转染人肺癌细胞株 A549。实时荧光定量PCR、Western blot检测KDR基因水平变化;流式细胞仪、MTT 法、集落形成试验检测其对A549细胞株细胞凋亡、细胞增殖和克隆形成的影响;细胞黏附实验评价其肿瘤靶向性。其抗癌作用主要表现为可有效地抑制A549细胞KDR基因表达、细胞增殖和克隆形成,促进细胞凋亡,并具有肿瘤靶向性作用。因而认为,TAT与KDR靶向siRNA慢病毒载体具有显著的肿瘤靶向性和抗肿瘤活性。