RNA干扰组蛋白去乙酰化酶1对人胰腺癌细胞增殖、凋亡的调控机制研究

目的观察沉默组蛋白去乙酰化酶1(HDAC1)基因对人胰腺癌PaTu8988细胞增殖、凋亡的影响及可能机制。方法胰腺癌PaTu8988细胞株分为空白对照组(不予任何处理)、阴性对照siRNA组(予以30nmol/L阴性siRNA)、siRNA1组(予以15nmol/LHDAC1siRNA)和siRNA2组(予以30nmol/LHDAC1siRNA)。siRNA转染48h后,用相对实时定量RT-PCR法和West-ernblotting检测HDAC1基因表达情况;相对实时定量RT-PCR法检测细胞增殖凋亡相关基因p21、Bcl-2、Bax的表达;WST-8法检测细胞增殖情况;流式细胞术检测细胞凋亡变化。结果转染HDAC1siRNA48h后人胰腺癌PaTu8988细胞中HDAC1mRNA表达量在siRNA1组和siRNA2组分别为46.1%±6.1%和32.3%±1.4%,均显著低于空白对照组(100.0%±3.4%)和阴性对照siRNA组(87.4%±28.3%,P<0.05)。Westernblotting显示HDAC1蛋白表达量亦明显下降,以siRNA2组表达量最低(P<0.05)。WST-8法检测显示4组细...     

Survivin siRNA联合紫杉醇对肺癌细胞株的生长抑制作用

目的探讨survivin siRNA对肺癌A549细胞化疗敏感性的影响。方法将A549细胞分为siRNA转染组、N-siRNA转染组、阴性对照组、空白对照组,用MTT法检测不同浓度紫杉醇联合siRNA对细胞存活率的影响。将A549细胞分为阴性对照组、siRNA组、紫杉醇组、siRNA 紫杉醇组,采用MTT法检测siRNA联合紫杉醇作用不同时间对细胞存活率的影响;采用Western blot检测细胞内survivin、p21、PARP蛋白表达变化;采用流式细胞仪分析各组细胞周期和凋亡变化的差异。结果Survivin siRNA可增加A549细胞对不同浓度紫杉醇的敏感性,但以低浓度(0.1、1、10nmol/L)最显著(P<0.05);survivin siRNA与10nmol/L紫杉醇共同处理A549细胞48h后,对细胞增殖的抑制具有协同或相加的效果(q值分别为1.34、1.03、1.02)。siRNA不仅可以抑制A549细胞内源性survivin表达还可以抑制紫杉醇诱导的survivin表达,阴性对照组、紫杉醇组、siRNA组和siRNA 紫杉醇组中survivin蛋白条带相对强度分别为0.244、0.99、0、0;survivin siRNA和紫杉醇联合作用后,对凋亡的诱导也有轻度相加作用,阴性对照组、紫杉醇组、siR-NA组和siRNA 紫杉醇组凋亡率分别为0.47%、9.50%、4.78%、19.81%;与对照组相比,siRNA和紫杉醇联合作用对细胞周期的影响表现为G1期、G2/M期细胞增加,S期细胞比率明显减少,对PARP的裂解和p21蛋白的表达也有促进作用。结论Survivin过表达可能参与了紫杉醇获得性耐药的形成,siRNA阻抑survivin表达是增加肺癌细胞对紫杉醇敏感性的有效途径。     

p53调控信号转导子和转录激活子3对肺癌细胞增殖及凋亡的影响

 目的 探讨p53调控信号转导子和转录激活子3(STAT3)对肺癌细胞增殖及凋亡的影响。方法 细胞分组为空白对照组、阴性对照组、p53 小干扰RNA(siRNA)组、p53 siRNA+STAT3 siRNA组,采用western blot法检测组织中p53和STAT3蛋白表达水平;采用Transwell法检测转染后体外迁移能力和侵袭能力;采用流式细胞术膜联蛋白Ⅴ/碘化丙啶(AV-PI)双染法检测转染后凋亡情况。结果 肺癌组织中p53表达水平明显下调,而STAT3表达阳性率增高,差异均有统计学意义(P<0.05)。转染p53的siRNA能显著上调肺癌细胞中STAT3的表达水平;p53 siRNA组和p53 siRNA+STAT3 siRNA组A549细胞凋亡率(17.91%±0.67%,22.51%±0.56%)、细胞的迁移速度[(55.16±6.50)μm/h,(63.30±5.98)μm/h]、穿膜细胞数(58.23±7.12,68.23±7.14),与空白对照组和阴性对照组比较,差异均有统计学意义(P<0.05)。结论 p53在肺癌中呈现低表达,p53能负向调控STAT3的表达,从而抑制其增殖和凋亡能力。     

应用RNAi技术沉默survivin基因对肺腺癌A549细胞的影响

目的 探讨RNA干扰(RNAi)沉默survivin基因对肺腺癌A549细胞的影响.方法 应用RNAi技术,以化学合成的survivin小干扰RNA(siRNA)体外转染A549细胞,采用MTT法及流式细胞仪检测转染前后A549细胞增殖、凋亡和细胞周期的变化,通过RT-PCR及Western blot分析survivin mRNA和蛋白的表达情况.结果 survivin siRNA对A549的生长抑制作用呈浓度依赖性(P＜0.05),并能引起G1期细胞比率的增加和S期细胞相应的减少,诱导一定数量的细胞凋亡,转染后36、60、84、108、132h的IC50值分别为152.4、151.2、136.0、144.0、150.4nmol/L.100～200nmol/L siRNA转染组细胞的survivin表达在蛋白及RNA水平都显著低于对照组(P＜0.01),但空白对照组与空质粒组间无明显差异(P＞0.05).结论 应用siRNA沉默survivin基因可以下调肺腺癌A549细胞survivin的表达,进而抑制细胞生长、增殖并诱导细胞凋亡.RNAi的抑制效果具有特异、高效和持久的特点.     

慢病毒载体介导HIF-1α RNAi对人胰腺癌细胞Patu8988相关基因表达的影响

目的 探讨慢病毒表达载体介导HIF-1α RNA干扰(RNAi)对人胰腺癌细胞Patu8988HIF-1α和Glut-1表达的影响.方法 构建针对HIF-1α基因的RNAi慢病毒表达载体LV-RNAi-HIF-1α.乏氧条件下体外培养4h,转染LV-RNAi-HIF-1α的Patu8988细胞为实验组;转染空病毒载体和未转染任何病毒载体Patu8988细胞分别为阴性对照组和空白对照组.采用荧光定量RT-PCR和Western印迹法检测Patu8988细胞HIF-1α表达情况,采用RT-PCR法检测转染LV-RNAi-HIF-1α后Patu8988细胞Glut-1的表达情况.采用SPSS 17.0软件行单因素方差分析和两样本t检验分析各组之间的差异.结果 构建的慢病毒表达载体LV-RNAi-HIF-1α,在常氧和乏氧状态下致HIF-1α mRNA表达分别下降65.1％(0.209/0.321)和80.6％ (0.791/0.982)(t=10.52和15.24,均P＜0.05),阴性对照组分别为0.6％(0.002/0.321)和7.2％(0.071/0.982)(t=5.26和7.38,均P＜0.05);乏氧状态下实验组、阴性对照组和空白对照组HIF-1α蛋白的表达分别为0.159±0.010、0.745±0.012和0.711±0.023,差异有统计学意义(F=35.52,t=6.72和10.56,均P＜0.05),实验组较其他2组表达下降;乏氧条件下实验组Glut-1 mRNA表达(0.040±0.003)较阴性对照组(0.054±0.003)和空白对照组(0.062±0.004)均有明显下降(F=35.28,t=5.94和8.55,均P＜0.01).结论 通过构建LV-RNAi-HIF-1α慢病毒表达载体沉默HIF-1α基因表达,可降低Patu8988胰腺癌细胞Glut-1 mRNA表达.     

DNA甲基转移酶基因干扰对HepG2细胞癌-睾丸抗原再表达的影响

目的 探讨抑制甲基转移酶(DNMT1、DNMT3a和DNMT3b)对肝细胞癌细胞中癌-睾丸抗原表达的影响及相关机制.方法 分别采用针对DNMT1、DNMT3a和DNMT3b的siRNA(DNMT1+3a+3b),针对DNMT1和DNMT3a的siRNA(DNMT1+3a),针对DNMT1和DNMT3b的siRNA(DNMTl+3b)以及针对DNMT3a和DNMT3b的siRNA(DNMT3a+3b)转染HepG2细胞,并以使用5-杂氮脱氧胞嘧啶(5-Aza-dC)处理的细胞作为阳性对照,采用RT-PCR、实时定量PCR和Western blotting检测细胞中DN-MT及癌-睾丸抗原的表达情况,并采用甲基化特异PCR(MSP)检测部分癌-睾丸抗原基因启动子的甲基化状态.结果 经siRNA干扰后,细胞中DNMT1、DNMT3a和DNMT3b的表达量均明显降低.癌-睾丸抗原CT10和SSX1在转染DNMT1+3a和DNMT1+3b的细胞中出现再表达,而MAGE1及MAGE3在各siRNA转染细胞中均有表达,且无明显差异.CT10的启动子区发生了去甲基化,但MAGE1启动子区处于非甲基化状态.结论 在HepG2细胞中,干扰DNMT可使癌-睾丸抗原基因的启动子区发生去甲基化,后者可使由甲基化导致的不能表达的癌-睾丸抗原基因发生再表达.     

RNA干涉抑制肺癌细胞DNA甲基转移酶1(DNMT1)的表达

目的：通过抑制肺癌细胞DNA甲基转移酶1(DNMT1)的表达，从而部分恢复抑癌基因的表达。方法：采用RNAi技术来抑制DNMT1的活性。应用体外转录的方法，共合成了5个针对DNMT1不同靶位的siRNA序列，并转染人肺癌细胞NCI-H157。采用MSP-PCR、COBRA等方法对抑癌基因RASSF1A进行甲基化分析，半定量RT-PCR检测DNMT1 mRNA的敲降与RASSF1A基因的表达。结果与结论：5个靶位的siRNA序列中有两个靶位能够有效地抑制DNMT1的表达，且呈剂量依赖效应。人肺癌细胞NCI-H157中抑癌基因RASSF1A发生了高度甲基化，在抑制DNMT1活性后，RASSF1A基因去甲基化而恢复表达。     

siRNA对骨肉瘤细胞细胞周期及survivin表达的影响

目的 构建人survivin特异性siRNA,探讨其对骨肉瘤细胞MG63细胞周期及survivin表达的影响.方法 构建survivin特异性的RNA干涉载体,转染MG63细胞,G418筛选稳定转染的细胞系,以Western blot法检测survivin蛋白表达变化,透射电镜、Hoechst染色检测细胞凋亡情况,流式细胞仪检测细胞周期变化.结果 成功构建了survivin基因siRNA真核表达载体PsiS,获得了稳定转染的细胞系.与正常MG63细胞、阴性对照细胞相比,MG63/PsiS细胞中survivin蛋白表达减少,部分细胞核致密浓染或碎块状浓染,凋亡率增加了7倍,而G2/M期细胞减少近50%.结论 特异性siRNA能够明显抑制survivin基因在MG63细胞中的表达,抑制细胞增殖,促进细胞凋亡.     

小干扰RNA抑制核因子-κB增强131I治疗甲状腺癌疗效的研究

目的 探讨小干扰RNA (siRNA)抑制核因子-κB (NF-κB)对131I致DTC细胞凋亡能否产生协同作用.方法 2×104 MBq/L 131I作用人甲状腺乳头状癌细胞株KTC-1 24 h后做DNA结合实验,48 h后做细胞存活分析.Western blot鉴定131I作用6h后细胞NF-κB p65的变化,24 h后凋亡抑制因子[X-染色体相关凋亡抑制蛋白(XIAP)、细胞凋亡抑制因子1(clAP1)、B细胞淋巴瘤因子大亚基(Bcl-xL)]、凋亡关键因子[半胱氨酸蛋白酶(caspase 3)和多聚腺苷二磷酸核糖聚合酶(PARP)]的变化.p65和凋亡抑制因子的Western blot检测分4组:未转染(A)组、未转染+131I(B)组、转染对照siRNA+ 131I(C)组和转染p65 siRNA+131I(D)组;其余实验分为6组:未转染(1)组、转染对照siRNA(2)组、转染p65 siRNA(3)组、未转染+131I(4)组、转染对照siRNA+131I(5)组和转染p65 siRNA+131I(6)组.多组间均数比较采用单因素方差分析,均数两两比较采用q检验.结果 1至6组DNA结合率分别为(100.00±11.65)％、(96.00±17.98)％、(9.28±5.01)％、(322.72±50.81)％、(311.36±44.81)％和(36.96±15.66)％,差异有统计学意义(F=137.74,P＜0.01);131I作用后KTC-1细胞NF-κB活性均增强(q4∶1组=10.90,q5∶2组=11.38,均P＜0.01);p65 siRNA可抑制NF-κB功能(q1∶3组=18.25,q4∶6组=13.71,均P＜0.01).6组细胞存活率分别为(100.00±11.65)％、(96.32±9.44)％、(70.88±7.41)％、(64.16±9.50)％、(62.24±9.37)％和(28.64±6.74)％ (F=52.76,P＜0.01);3、4和6组比,q=10.76和7.79,均P＜0.01.Western blot结果显示A、B、C和D组p65相对表达水平分别为(56.60 ±7.37)％、(111.07±13.31)％、(113.16±15.04)％和(12.46±2.74)％,差异有统计学意义(F=60.17,P＜0.01);131I作用后p65浓度增高(qB∶A组=6.20,qc∶A组=5.85,均P＜0.01);p65 siRNA可抑制其浓度增高(qB∶D组=12.57,qc∶D组=11.41,均P＜0.01).4组XIAP、cIAP1和Bcl-xL分别为(17.59±1.96)％、(16.45±1.85)％和(19.92±2.22)％,(98.37±17.92)％、(109.81±19.16)％和(95.59±22.20)％,(98.43±18.71)％、(98.86±15.88)％和(100.99±21.70)％,(7.00±0.95)％、(5.86±0.35)％和(9.52±0.90)％,差异均有统计学意义(F =44.22、56.51和29.11,均P＜0.01);131I作用后三者表达增加(qB∶A组 =7.76、8.40和5.88,均P＜0.01);p65 siRNA可抑制三者表达(qB∶D组=8.82、9.40和6.71,均P＜0.01).6组caspase 3亚基p19和p17、PARP活性蛋白p116和失活产物p89差异均有统计学意义(F=39.03、48.45、32.56和52.20,均P＜0.01);3、4和6组比q =3.18 ～9.98,均P＜0.05.结论 131I通过活化NF-κB导致甲状腺癌细胞内凋亡抑制因子表达升高,p65 siRNA可抑制这种变化;联合使用p65 siRNA对131I致DTC细胞凋亡产生协同效应.     

miR-451对结肠癌细胞系SW620生物学行为的影响

目的探讨miR-451对结直肠癌细胞SW620增殖、凋亡及侵袭能力的影响。方法将SW620细胞分为4组:miR-451模拟物(miR-451 mi mics)转染组(细胞转染miR-451 mi mics,终浓度100nmol/L),NC组(细胞转染miRNA阴性对照,终浓度100nmol/L),空白转染组(加入与前2组等量的Lipofectamine 2000,无miRNA片段),对照组(细胞常规培养,不加入Lipofectamine 2000和miR-NA片段)。应用荧光定量RT-PCR检测转染24h后miR-451表达量的改变,采用CCK-8试剂盒检测转染24、48、72h后细胞的增殖能力,流式细胞仪检测转染48h后细胞凋亡情况,Transwell侵袭实验检测转染24h后细胞侵袭能力的改变,细胞免疫荧光和Westernblotting检测转染48h后巨噬细胞迁移抑制因子(MIF)的表达情况。结果转染后24h,荧光定量RT-PCR检测结果显示,miR-451mi mics转染组的miR-451表达量(67.96±13.33)较NC组(以其miR-451表达量作为1)显著上调(P<0.01)。与另外3组比较,miR-451 mi mics转染组细胞增殖能力在转染后48、72h明显受抑(P<0.01),但转染后48h其细胞凋亡率无明显改变(P>0.05)。转染后48h,miR-451 mi mics转染组、NC组、空白转染组、对照组中MIF蛋白表达阳性细胞的比例分别为23.9%±14.9%、53.5%±14.1%、45.2%±19.8%、59.3%±22.2%,统计学分析显示miR-451 mi mics转染组与其他3组比较差异显著(F=7.356,P<0.01)。Transwell侵袭实验显示,miR-451 mi mics转染组侵袭细胞数(37.4±6.1个/视野)明显低于对照组(61.6±8.6个/视野)、NC组(55.6±3.6个/视野)、空白转染组(60.8±7.4个/视野,P均<0.01)。结论上调miR-451表达可抑制结直肠癌细胞的生物活性,该作用可能与抑制MIF表达有关。     

Fluorodeoxyglucose cell incorporation as an index of cell proliferation: evaluation of accuracy in cell culture

The use of fluorodeoxyglucose (FDG) and positron emission tomography (PET) is recognized as an accurate tool for the specific diagnosis and staging of cancer. It has also been proposed for the monitoring of anticancer therapy. FDG cell incorporation reflects glycolytic activity whereas inhibition of cell proliferation corresponds to an efficient cancer treatment. The relationship between FDG incorporation and cell proliferation has yet to be demonstrated. Therefore, we aimed to correlate the effects of the toxic agents bleomycin and unlabelled meta-iodobenzylguanidine (mIBG) on cellular metabolism and proliferation. We determined the in vitro metabolic and cytotoxic effects of bleomycin and mIBG by measuring the incorporation of fluorine-18 FDG (%U_FDG) and hydrogen-3 thymidine (%U_THY) in cells of the human premonocytic line U937 in the presence of increasing concentrations of these agents. Proliferation rate of these cells was studied by means of limiting dilution analysis. %U_THY appeared more sensitive to bleomycin or mIBG-mediated cell injury than %U_FDG. After 1 h of exposure to 0.5 M bleomycin, %U_THY was significantly reduced to 62.0% ± 10.4% of control value whereas %U_FDG remained unchanged (91.6% ± 5.3%). Similar results were obtained after 1 h of exposure to increasing concentrations of mIBG (1 M to 1 mM). After 20 h of exposure to bleomycin, %U_THY and %U_FDG were significantly reduced as a function of concentration. After 20 h of exposure to mIBG, a transient increase in %U_FDG up to 149.3% ± 11.2% with 50 M mIBG was further followed by a reduction to 20.1% ± 6.7% with 0.5 mM (P < 0.001). The clonogenic efficiency was reduced as a function of bleomycin (ANOVA, n=255, P) or mIBG concentration (n=80, P) and nearly abolished with 0.1 M bleomycin or 0.1 mM mIBG. In conclusion, %U_THY appears to be a more sensitive index of cytotoxicity in vitro and more accurately relates to cell proliferation than %U_FDG. Correspondence to: D.O. Slosman, Nuclear Medicine Division, Geneva University Hospital, CH-1211 Geneva 14, Switzerland     

透明细胞乳头状肾细胞癌的影像学表现

目的 探讨透明细胞乳头状肾细胞癌(CCPRCC)的影像学表现.方法 分析15例CCPRCC患者CT及MRI影像特征,采用独立样本t检验比较肿瘤与肾皮质之间平扫CT值、ADC值差异.结果 15例均为单发,边界清晰,大小为(3.1±1.9) cm.13例为实性肿瘤,其中11例伴囊变,2例为囊性肿瘤.4例CT平扫呈等或稍低密...     

肾透明细胞癌与肾乳头状癌、嫌色细胞癌MRI表现的对照研究

目的 探讨MR动态增强扫描对肾癌亚型的鉴别诊断价值.方法 搜集77例经病理证实的肾癌患者资料,其中透明细胞癌(CCRCC)55例,乳头状癌(PRCC)14例,嫌色细胞癌(CRCC)8例,回顾性分析各亚型肿瘤患者MR平扫及动态增强扫描表现并与病理对照,根据肿瘤及肾皮质增强前后的皮质期、实质期及延迟期信号变化,分别进行百分比测量、肿瘤-肾皮质增强指数计算,并采用单因素方差分析和LSD法进行比较.结果 CRCC多数信号均匀(7/8);CCRCC及PRCC多数信号不均(分别为51/55和13/14)、常见坏死(36/55和7/14),PRCC最常见出血(9/14)及囊变(9/14).动态增强各期CCRCC强化程度最高,强化模式呈"快进快退",CRCC轻至中度强化,PRCC强化最轻,两者均呈渐进性延迟强化.CCRCC、PRCC及CRCC皮质期信号变化分别为(296.15±60.27)%、(79.70±18.84)%和(119.56±40.76)%,实质期分别为(236.33±58.31)%、(122.81±27.35)%和(163.06±33.91)%,延迟期分别为(216.83±46.72)%、(117.55±20.63)%和(179.72±32.89)%;三者皮质期的肿瘤-皮质增强指数分别为1.26±0.34、0.33±0.12及0.54±0.10,实质期分别为0.92±0.23、0.41±0.23及0.62±0.15,延迟期分别为0.76±0.14、0.35±0.11及0.69±0.12,各亚型增强各期的信号变化(F值分别为940.931、124.515、38.194,P值均＜0.01)、肿瘤-皮质增强指数(F值分别为798.625、78.308、73.699,P值均＜0.01)差异均有统计学意义.3种亚型的MRI表现与病理学所见基本相符.结论 CCRCC、PRCC及CRCC的MRI动态增强有一定特征性的表现,与其病理特点密切相关,在肾癌亚型的鉴别诊断上有着较高的临床应用价值.

Abstract：

Objective To investigate the differential diagnostic features of subtypes of renal cell carcinoma(RCC) using dynamic contrast-enhanced MRI(DCE-MRI).Methods The MRI appearances of 77 RCCs, including 55 clear cell RCCs(CCRCC),14 papillary RCCs(PRCC) and 8 chromophobe RCCs(CRCC), were retrospectively analyzed and compared with findings of pathology. DCE-MRI was conducted in each case after intravenous administration of contrast agent. Region of interest measurements (cortical, nephrographic and delayed Phases) of signals within tumor and uninvolved renal cortex were used to calculate percentage signal intensity change and tumor-to-cortex enhancement index, and the data was analyzed by AVONA and t test. Results On unenhanced and enhanced MRI, most CRCCs showed homogeneous signal(7/8). CCRCC and PRCC often show inhomogenous signal with necrosis(36/55, 7/14). Hemorrhage and cystic degeneration were often found in PRCC (9/14). On the cortical, nephrographic and delayed phase images, CCRCCs showed greater signal intensity change[(296.15±60.27)%, (236.33±58.31)% and (216.83±46.72)%,respectively than PRCCs (79.70±18.84)%, (122.81±27.35)% and (117.55±20.63)%, respectively], and CRCCs showed intermediate change [(119.56±40.76)%, (163.06±33.91)% and (179.72±32.89)%, respectively].A phenomenon of quick staining and quick fainting was observed in CCRCCs. Both of CRCCs and PRCCs showed delayed enhancement. The tumor-to-cortex enhancement index at the cortical, nephrographic and delayed phases was highest for CCRCCs (1.26±0.34, 0.92±0.23 and 0.76±0.14, respectively), lowest for PRCCs (0.33±0.12, 0.41±0.23 and 0.35±0.11, respectively), and intermediate for CRCCs (0.54±0.10, 0.62±0.15 and 0.69±0.12, respectively,P＜0.01). The degree of enhancement was significantly different among the 3 subtypes at the every contrast enhanced phase (F=940.931, 124.515 and 38.194, P＜0.01), so was the tumor-to-cortex enhancement index(F=798.625,78.308 and 73.699, P＜0.01). There was a good consistency between MR appearances of the 3 RCC subtypes and pathological characteristics. Conclusion DCE-MRI could distinctly show imaging features of CCRCC, PRCC and CRCC, which were related to their pathological characteristics, and these features were helpful in predicting a specific subtype of RCC.     

一种改进的用于测定细胞周期的细胞制备方法

目的 :减少细胞的聚集数量 ,提高测试效率和结果的准确性。方法 :在用酒精固定细胞时分别加入终浓度为 0 % ,1.5 % ,3% ,6 %和 12 %的小牛血清 ,置 - 2 0℃分别固定细胞 1d ,3d和 7d。比较了不同浓度的血清和保存不同时间粘连细胞的数量及对细胞周期分析结果的影响。结果 :加入血清可明显减轻细胞的粘连 ,减少了细胞的聚集数量 ,尤以 3%小牛血清组最佳。样品可不必过滤 ,在上机测试时 ,进样针不堵塞 ,上样速度快 ,细胞周期分析更准确。随着保存时间的延长 ,聚集细胞的数量有增加的趋势。结论 :在制备用于测定细胞周期的样品时 ,固定细胞的过程中加入终浓度为 3%的小牛血清是一种简单的、能有效地保护细胞膜使细胞不易粘连的技术措施 ,且固定细胞的时间不宜超过 7d。     

电离辐射对不同肿瘤细胞细胞周期的影响

目的 研究电离辐射对不同肿瘤细胞细胞周期的影响为肿瘤放疗及化疗提供科学依据。方法 处于细胞周期各时相的细胞百分数采用流式细胞术进行检测。结果 研究表明：电离辐射作用后,ＨｅｌａＳ３和Ｓ１８０细胞发生了Ｓ和Ｇ２期阻滞,而ＤＬ－４细胞则发生Ｇ１和Ｇ２期阻滞,Ｂ１６各时相细胞数无列出较高的辐射抗性。结论 电离辐射作用后,不同肿瘤细胞的辐射抗性、即辐射敏感性有较大差异。其细胞周期的变化规律亦不相同。     

胶质瘤干细胞的生存微环境及其示踪研究进展

胶质瘤干细胞在胶质瘤的形成和生长中发挥至关重要的作用。胶质瘤干细胞的生存微环境是维持干细胞特性的关键,在体无创性示踪为研究胶质瘤干细胞的生存、繁殖、自我更新等特性以及促进胶质瘤发展的机制具有重要意义。     

Potency of different red light sources in photodynamic induction of cell death in a squamous cell carcinoma cell line

LED illumination systems were found to be more efficacious than broad spectrum lamps in recent phase III trials on photodynamic treatment of actinic keratosis. However, a detailed comparison of the light doses emitted at the appropriate spectral range and its correlation to photodynamic effects is thus far not available for the most frequently used devices. Here, we compared the spectral emissions of three different PDT lamps with their potency of inducing cell death in ALA-loaded A431 cells, including a new system equipped with more advanced LEDs matching the photosensitizer absorption peak more precisely and emitting more homogeneous light over time. Cells were exposed to two different ALA concentrations, incubated for 1 or 3 h and then illuminated by one of two different LED or a broad-spectrum system at four different light doses, whereupon viability was assessed. Maximal doses were selected in accordance to clinically applied light doses in recent phase III studies and the manufacturers’ recommendations. The data gathered here clearly demonstrate that the two LED systems were significantly more effective in inducing cell death than the broad spectrum system. Most efficient was the newer LED system, in agreement with emission parameters that more accurately corresponded to the photosensitizer’s absorption peak.     

敲低PES1基因抑制舌癌细胞的生长

目的：构建稳定表达PES1 shRNA的舌癌细胞系,研究敲低PES1基因对舌癌细胞生长的影响。方法在293 T细胞中包装并获得敲低PES1基因的病毒,然后将病毒感染Tca8113、SCC6和SCC153种舌癌细胞并筛选稳定克隆,Western印迹检测PES1蛋白的表达水平;利用生长曲线检测敲低PES1对舌癌细胞生长的影响,流式细胞术检测敲低PES1对舌癌细胞周期的影响。结果成功构建稳定表达PES1 shRNA的舌癌细胞系,敲低PES1基因能够抑制舌癌细胞的生长,使细胞周期阻滞在G0／G1期,敲低PES1抑制cyclin D1的表达。结论敲低PES1抑制舌癌细胞的生长,诱导细胞周期阻滞在G0／G1期,抑制cyclin D1的表达,因此PES1可能成为舌癌基因治疗的靶标。     

Synchronous ipsilateral renal cell and transitional cell carcinoma

The coexistence of multiple and synchronous primary neoplasms in the genitourinary system has only rarely been described in the literature. We present the case of a 78-year-old man with haematuria as the initial presentation, finally proven to be transitional cell carcinoma (TCC) combined with renal cell carcinoma (RCC). Intravenous urography (IVU), CT and arterial angiography studies revealed a space-occupying nodule at the right upper renal pelvicalyces showing mild enhancement with contrast medium. Another strong contrast medium enhancing exophytic tumour was found at the lower pole of kidney; there were hypodense foci and calcified components in this lesion. A right nephroureterectomy was performed. Pathological diagnosis was a papillary TCC and a clear cell type RCC. This is a rare case of combined renal malignancies diagnosed by imaging.