非小细胞肺癌组织中P27表达及定位与磷酸化AKT的关系

背景与目的:抑癌基因的失活在肿瘤的发生发展中有重要作用。p27的失活包括P27表达的减少或丢失及P27的胞浆错定位。AKT是一种丝氨酸/苏氨酸激酶,活化状态的AKT[磷酸化AKT(phosphorylatedAKT),p-AKT]能够调节细胞周期中的多个效应分子,其中包括对P27表达及定位的调节。本研究旨在探讨P27在非小细胞肺癌中(NSCLC)中的表达、定位及与p-AKT之间的相关性。方法:应用免疫组化的方法,分别检测NSCLC组织80例、非肿瘤性肺组织标本35例中P27蛋白的核、浆表达及磷酸化AKT的表达,并对它们与临床病理因素的相关性进行分析。结果:在NSCLC组织中,P27蛋白总阳性率为72.5%,其中胞核阳性率为46.3%;在非肿瘤性肺组织中P27总阳性率及胞核阳性率均为94.3%,两组对比P27总阳性率及胞核阳性率差异均有显著性意义(P<0.01)。80例NSCLC中有21例呈现单纯胞浆阳性;而在非肿瘤性肺组织中未见单纯胞浆阳性表达,两组对比差异有显著性意义(P<0.01)。P27的胞核表达在无淋巴结转移组和病理高、中分化组明显高于淋巴结转移组、低分化组的表达,组间比较差别具有显著性意义(P<0.05);P27的胞核表达与患者年龄、性别、肿瘤类型、TNM分期无关(P>0.05)。P27的胞浆表达与临床病理特征无关(P>0.05)。p-AKT蛋白在NSCLC中阳性表达率为78.8%,在非肿瘤性肺组织中呈阴性表达,两组之间差异有显著性意义(P<0.05);p-AKT在腺癌高、中分化组的表达(95.0%)高于低分化组的表达(50.0%)(P<0.01)。在NSCLC中,p-AKT的表达与P27胞浆表达呈正相关(r=0.437,P<0.01);p-AKT的表达与P27胞核表达无相关关系(r=0.175,P>0.05)。结论:NSCLCP27胞核表达的减少或丢失可能与肺癌的恶性发展有关;NSCLC中P27单纯性胞浆表达可能是肺癌细胞表达的特征之一;磷酸化AKT可能参与了对P27胞浆定位的调节,但与P27胞核表达无明显相关关系。     

E-cadherin、CD44v6和PCNA在非小细胞肺癌组织中的表达及意义

背景与目的:上皮钙依赖粘附蛋白(E-cadherin,E-cad)、CD44v6和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在恶性肿瘤侵袭与转移中发挥重要作用。本研究旨在探讨E-cad、CD44v6和PCNA在非小细胞肺癌(non-small cell lung cancer,NSCLC)及癌旁组织中的表达与NSCLC的侵袭、转移及预后的关系。方法:应用免疫组化EnVision法检测86例NSCLC组织及40例癌旁组织中E-cad、CD44v6和PCNA的表达,并分析与NSCLC的侵袭、转移及预后的关系。结果:E-cad在肺癌组织中的表达率为53.5%(46/86),显著低于癌旁组织(80.0%)(P<0.05),且与NSCLC的分化程度、淋巴结转移和TNM分期有关(P<0.05);CD44v6在肺癌组织中的表达率为44.2%(38/86),在癌旁组织中无表达,且在鳞癌中的表达率(54.0%)显著高于腺癌(30.6%)(P<0.05),并且与淋巴结转移和TNM分期有关(P<0.05);PCNA在肺癌组织中的表达率为48.8%(42/86),在癌旁组织中无表达,且在淋巴结转移组与未转移组间的表达差异有统计学意义(P<0.05)。E-cad与PCNA的表达呈显著性负相关(r=-0.554,P<0.05),而CD44v6与PCNA的表达呈显著性正相关(r=0.688,P<0.05)。单因素生存分析显示E-cad、CD44v6及PCNA的表达与NSCLC患者预后有关。Cox比例风险模型进行多因素生存分析显示;E-cad与临床分期是有意义的预后指标(P<0.05)。结论:E-cad、CD44v6及PCNA的表达与NSCLC的侵袭和转移相关。在NSCLC中联合检测三者的表达对判断预后有参考价值。     

线粒体融合蛋白-2在非小细胞肺癌组织中的表达及其临床意义

目的:探讨线粒体融合蛋白2(mitofusin 2,Mfn2)在非小细胞肺癌(non-small-cell lung cancer,NSCLC)组织和非癌肺组织中的表达及与临床病理特征之间的关系.方法:应用原位杂交和免疫组织化学法检测92例NSCLC组织和27例非癌肺组织中Mfn2 mRNA及蛋白的表达.结果: 肺鳞癌组织中Mfn2 mRNA和蛋白的阳性表达率(83.33%和89.58%)均高于肺腺癌(56.82%和65.91%),且2者均高于非癌肺组织的阳性表达率(25.93%和29.63%),3组间比较差异有统计学意义(P<0.001和 P<0.01).高分化和中分化的肺癌组织中Mfn2 mRNA和蛋白的阳性表达率(93.75%和100%,91.67%和91.67%)均高于低分化的肺癌组织(21.43%和42.86%),差异有统计学意义(P<0.001).Mfn2 mRNA和蛋白的表达与患者的性别、年龄、肿瘤大小、TNM分期和淋巴结转移无关(P>0.05).肺癌组织中Mfn2 mRNA和蛋白的表达具有较好的一致性.结论:Mfn2参与了肺癌的发生和发展,Mfn2的表达与肺癌的组织学类型及分化程度有关.     

非小细胞肺癌组织中MT蛋白与PCNA表达的研究

目的:研究金属硫蛋白(metallothionein, MT)和增殖细胞核抗原(proliferatingcellnuclearan tigen,PCNA)在非小细胞肺癌(non smallcelllung carcinoma,NSCLC)中的表达及其与临床病理因素 的关系。方法:选取NSCLC组织62例(鳞癌38 例,腺癌24例)及正常肺支气管黏膜组织15例石 蜡标本,用SP法检测MT及PCNA在各组织中的表 达。结果:MT蛋白和PCNA在NSCLC中的阳性表 达率分别为53.2%(33/62)和79.0%(49/62),均 明显高于正常肺支气管黏膜组织[20.0%(3/15)和 46.7%(7/15)],差异有统计学意义,P<0.05。肺 鳞癌和腺癌中MT的阳性表达率分别为50.0% (19/38)和58.3%(14/24);PCNA的阳性表达率分 别为76.3%(29/38)和83.3(20/24),其差异均无 统计学意义。MT蛋白和PCNA的阳性表达率均与 NSCLC组织的分化程度、淋巴结转移等因素密切相 关,差异有统计学意义,P<0.05。MT与PCNA阳 性表达呈正相关,P=0.004。结论:MT蛋白的高 表达可能是NSCLC发生的一种生物学标志,且与 PCNA存在协同作用。联合检测两种蛋白的表达水 平对判断NSCLC恶性程度、淋巴转移趋势具有重要 价值。     

血管生成和p16、p53蛋白表达与非小细胞肺癌淋巴结转移的相关性研究

目的　分析非小细胞肺癌 (NSCLC)淋巴结转移与血管生成及p16、p5 3蛋白表达的关系。方法　采用免疫组织化学方法对 187例NSCLC(74例腺癌、76例鳞癌和 37例腺鳞混合癌 )进行血管染色计数和p16、p5 3蛋白测定 ,并作相关临床因素分析。结果　非小细胞肺癌组织中微血管密度平均为 6 .8± 3.3,p16和p5 3蛋白阳性表达率分别为 4 7.6 %和 34.2 %。各组织类型非小细胞肺癌中淋巴结转移病例 (T2 N1～ 2 M0 )的微血管密度显著高于未转移者 (T2 N0 M0 ) ;在肺鳞癌和腺鳞癌中 ,p16和p5 3蛋白同时表达异常 (p16 -p5 3 )与微血管密度增高有关 ,但与淋巴结转移无关 ;而在肺腺癌中 ,p16和p5 3蛋白同时表达异常与淋巴结转移有关 ,但与微血管密度无关。微血管密度和p16、p5 3蛋白表达异常与年龄、性别、病理类型及分化程度均无明显相关。结论　各组织类型非小细胞肺癌的淋巴结转移均与活跃的血管生成密切相关 ,其中在肺鳞癌和腺鳞癌中 ,血管生成与p16和p5 3基因表达异常有关。而在肺腺癌中 ,血管生成和p16、p5 3蛋白异常表达是影响其淋巴结转移的两项独立的因素。     

基质金属蛋白酶在肺癌转移中作用的研究

目的:观察基质金属蛋白酶MMP2和MMP9在肺癌组织中的表达,探讨其与肺癌浸润及转移的关系。方法:采用免疫组织化学及逆转录聚合酶链反应(RTPCR)的方法分别检测30例非癌肺组织及有随访资料的87例肺癌组织中MMP2和MMP9及其mRNA的表达。结果:MMP2和MMP9在肺癌与非癌肺组织均存在阳性表达,但在肺癌组织表达率高于非癌肺组织(P<0.05),小细胞肺癌、鳞癌、腺癌和非癌肺组织MMP2阳性率分别为85.7%、44.7%、47.6%和23.3%;MMP9阳性率分别为78.6%、47.4%、38.1%和20.0%,其中小细胞肺癌组织中表达率最高(P<0.05)。MMP2和MMP9mRNA在低分化程度肺癌组织中表达较高,Ⅰ级最高,Ⅱ级次之,Ⅲ级最低;在不同肿瘤类型之间MMP2和MMP9mRNA表达有显著差异,小细胞肺癌的MMP2及MMP9mRNA表达量明显高于腺癌和鳞癌;肺癌伴淋巴结转移患者癌组织MMP2及MMP9mRNA表达量明显高于未转移者。结论:MMP2和MMP9的表达与肺癌的病理类型、分化程度及是否有淋巴结转移等因素有关,MMP2和MMP9可能在肺癌侵袭转移中起重要作用。     

COX-2和p53蛋白在非小细胞肺癌中的表达及与微血管生成的相关性研究

目的探讨COX-2、p53的表达与肺癌血管生成及临床病理特征的关系。方法收集临床资料完整的肺癌手术标本85例,肺良性病变手术标本20例,采用免疫组化方法检测COX-2、p53蛋白及MVD的表达情况。结果(1)COX-2、p53在肺癌中阳性表达率分别为56.5%和58.8%,肺癌MVD值为43.55±11.45,均显著高于肺良性病变(P<0.05);(2)肺癌组织中COX-2表达与组织学类型、肿瘤分化程度和淋巴结转移显著相关(P<0.05);(3)肺癌组织中p53表达与淋巴结转移相关(P<0.05);(4)肺腺癌MVD值显著高于肺鳞癌,有淋巴结转移组MVD值显著高于无淋巴结转移组(P<0.05);(5)COX-2与MVD、p53与MVD、COX-2与p53的表达呈正相关。结论COX-2、p53的表达增强参与了肺癌的发生发展过程,两者在肺癌血管生成方面存在协同作用。联合检测两者的表达情况对肺癌的临床评价有重要价值,并可能成为肺癌治疗的新靶点。     

MT1-MMP和MMP11在肺癌中的表达及其预后价值

目的探讨MT1-MMP和MMP11在肺癌组织中的表达及其与浸润转移和预后的关系。方法采用免疫组化检测47例肺鳞癌和42例肺腺癌组织中MT1-MMP和MMP11蛋白的表达。结果 MT1-MMP蛋白阳性表达率在肺鳞癌和腺癌组织中分别为68.1%(32/47)和64.3%(27/42)(P=0.705),其表达与T分期、淋巴结转移和TNM分期相关(P<0.05)。MMP11蛋白在肺鳞癌和腺癌组织中的阳性表达率分别为61.7%(29/47)和57.1%(24/42)(P=0.662),其表达与淋巴结转移和TNM分期相关(P<0.05)。MT1-MMP和MMP11表达呈正相关(γ=0.332,P=0.001)。Kaplan-Meier生存曲线显示MT1-MMP和MMP11阳性表达组5年生存率均显著低于阴性表达组(P<0.05)。结论MT1-MMP和MMP11的表达与肺癌的浸润转移密切相关,两者的阳性表达均提示肺癌患者的不良预后。     

缺氧诱导因子-1α表达与大肠腺癌细胞增殖及血管形成的关系

目的探讨缺氧诱导因子1α(HIF1α)在大肠腺癌组织中的表达及其与细胞增殖、血管形成的关系。方法应用免疫组织化学方法检测20例正常大肠组织、60例大肠腺癌组织中HIF1α、增殖细胞核抗原(PCNA)蛋白表达及微血管密度(MVD),并分析HIF1α表达与PCNA、MVD的相关性。结果HIF1α蛋白在大肠腺癌中的阳性表达率为73.3%(44/60),其阳性表达与大肠腺癌淋巴结转移、Dukes分期有关,其阳性表达率淋巴结转移组(89.3%,25/28)显著高于无淋巴结转移组(59.4%,19/32)(P<0.05),并随肿瘤分期的升高而增高。HIF1α表达与PCNA、MVD呈正相关(γ=0.647,P<0.01和γ=0.655,P<0.01)。结论HIF1α在大肠腺癌组织中表达水平明显上调,并与肿瘤细胞增殖、血管形成密切相关,提示它可能在大肠癌的发生发展过程中起重要作用。     

肺腺癌和鳞癌组织中mdm2蛋白表达的临床研究

目的了解mdm2蛋白的表达与肺腺癌、鳞癌临床病理特征之间的关系及意义,探讨mdm2蛋白在肺癌发生、发展过程中的可能作用。方法用免疫组化法(SP法)检测mdm2蛋白在53例肺癌组织和19例癌旁正常肺组织的表达情况,并结合肺癌的临床病理特征进行对比分析。结果mdm2蛋白定位于细胞质或细胞核,为棕黄色颗粒。53例肺癌组织mdm2蛋白过表达率为52.8% (28/53),19例癌旁正常肺组织均不表达mdm2蛋白,二者差异有显著性(P<0.05)。Ⅲ～Ⅳ期和有淋巴结转移肺癌的mdm2蛋白过表达率分别为71.4%(15/21)和67.7%(21/31),显著高于Ⅰ～Ⅱ期(40.6%)和无淋巴结转移的过表达率(31.8%,P<0.05)。低分化肺癌mdm2蛋白过表达率90.0% (9/10)显著高于高分化及中分化的过表达率(40.0%和45.5%,P<0.05)。mdm2蛋白过表达率与患者年龄、性别、吸烟史及病理类型无显著相关性(P>0.05)。mdm2蛋白过表达组肺癌患者的术后5年生存率显著低于不表达组(P<0.05)。结论mdm2蛋白在肺癌中过表达与淋巴结转移、pTNM分期、分化程度有关。mdm2蛋白过表达可能在肺癌的发生、发展及预后中起重要作用。     

人干扰素对HL60,K562细胞生长及细胞周期的影响

用人干扰素(α和γ)与HL60、K562细胞共同培养后,对细胞生长有不同程度抑制作用。IFN_r对细胞生长抑制作用强于IFN_r,IFN_r和IFN_r联合应用有协同作用,在K562细胞,细胞在细胞周期中的分布也发生改变,G_0/G_1期细胞比例减少,S期细胞比例增高,在HL60细胞则无明显的细胞周期再分布情况。提示细胞在S期的堆积是细胞生长受抑的原因之一。     

六种小鼠肿瘤及白血病细胞株的细胞动力学研究

本文对L7811-85,L7212-85,L1210-86,P388-86 4个淋巴细胞白血病及S180-86和肝Ca-86肿瘤细胞株分别测定了细胞生长曲线、克隆形成效率和细胞DNA含量分布等参数。对其中L7811-85细胞株进行了较系统的细胞动力学研究,并应用~3H-TdR自杀试验微培养测定其克隆形成细胞的增殖时和合成时。结果表明:这6种细胞株均可在体外无刺激因子作用下良好生长。并可见随细胞传代数增加,细胞增殖加速,克隆形成率提高。用~3H-TdR自杀试验测出的克隆形成细胞增殖时与群体细胞倍增时相比,可反映其潜在倍增时。     

树突状细胞肉瘤(DCS)细胞的建立及其在侵袭转移中的应用

利用LII移植瘤,我们建立的小鼠树突状细胞肉瘤(DCS)细胞系,经过不同时期不同方法(形态、免疫组化、表达基因组芯片)的不断验证[李存玺、zhou shixin],确立了其为DC细胞的起源.     

Downregulation of TMPRSS4 Enhances Triple-Negative Breast Cancer Cell Radiosensitivity Through Cell Cycle and Cell Apoptosis Process Impairment

Background: Radioresistance remains a challenge for cancer radiotherapy. The present study aims to investigate the role of TMPRSS4 in triple negative breast cancer (TNBC) cell radiosensitivity. Materials and Methods: After transfection of MDA-MD-468 triple negative breast cancer cells line by using the lentivirus vector, the effect of TMPRSS4 down-regulation on TNBC radiosensitivity was evaluated by using cloning assay and CCK-8 assay. The CCK-8 assay was also used for performing cell proliferation analysis. Western blot was carried out to detect the expression of certain proteins related to cell cycle pathways (cyclin D1), cell apoptosis pathways (Bax, Bcl2, and Caspase3), DNA damage and DNA damage repair (TRF2, Ku80 , ˠH2AX) . The cell cycle and cell apoptosis were also investigated using flow cytometer analysis. Results: TMPRSS4 expression was down-regulated in MDA-MB-468 cells which enhanced MDA-MB-468 cells radiosensitivity. TMPRSS4 silencing also improved IR induced cell proliferation ability reduction and promoted cell arrested at G2/M phase mediated by 6 Gy IR associated with cyclin D1 expression inhibition. Moreover, TMPRSS4 inhibition enhanced TNBC apoptosis induced by 6 Gy IR following by over-expression of (Bax, Caspase3) and down-regulation of Bcl2 as the pro-apoptotic and anti-apoptotic proteins, respectively. Otherwise, TMPRSS4 down-regulation increases  DNA damage induced by 6 Gy IR and delays DNA damage repair respectively illustrated by downregulation of TRF2 and permanent increase of Ku80 and ˠH2AX expression at 1 h and 10 h post-IR. Conclusion: Down-regulation of TMPRSS4 increases triple negative breast cancer cell radiosensitivity and the use of TMPRSS4 inhibitor can be encouraged for improving radiotherapy effectiveness in TNBC radioresistant patients.     

Enhancement of Fluoropyrimidine Inhibition of Cell Growth by Leucovorin and Deoxynucleosides in a Human Squamous Cell Carcinoma Cell Line

The growth inhibitory effects of the fluoropyrimidines 5-flourouracil (5-FU) and 5-fluoro-2'-deoxyuridine (FdUrd) against a human squamous cell carcinoma cell line (SQ-1) were studied in the absence and presence of deoxynucleosides and/or N⁵-formyl-tetrahydrofolate (leucovorin). Inhibition of cell growth by the fluoropyrimidines was less when undialyzed rather than dialyzed fetal bovine serum was used. Leucovorin, in concentrations of 10-^-6 to 10-^-4M potentiated the growth inhibition of FU and FdUrd; deoxyguanosine and deoxyinosine in concentrations of 10^-5M also enhanced the growth inhibition produced by these fluoropyrimidines. In the presence of leucovorin addition, deoxyguanosine (10-^-5M) caused a further synergistic inhibition of cell growth produced by FdUrd but not FU. In contrast, addition of deoxyinosine at 10-^-5M resulted in further potentiation of FU but not FdUrd inhibition of cell growth in the presence of leucovorin. The synergy obtained with these combinations encourage the exploration of modulation of fluoropyrimidine by leucovorin and deoxynucleosides in vivo.     

Silencing of lncRNA AFAP1-AS1 Inhibits Cell Growth and Metastasis in Clear Cell Renal Cell Carcinoma

The lncRNA AFAP1-AS1, oriented from an antisense direction to the protein-coding gene AFAP1 in the opposite strand, was upregulated in a variety of tumors and associated with poor prognosis, including lung cancer, breast cancer, ovarian cancer, and so on. However, the biological role of AFAP1-AS1 in clear cell renal cell carcinoma (ccRCC) is still unknown. We observed that AFAP1-AS1 expression was significantly upregulated in ccRCC tissues and that patients with high-level expression of AFAP1-AS1 had a shorter overall survival. Knockdown of AFAP1-AS1 markedly suppressed the progression of proliferation, invasion, migration, and EMT in ccRCC cells. Downregulation of AFAP1-AS1 resulted in an increase in E-cadherin and a decrease in vimentin. Noticeably, we found that PTEN has a negative correlation with the lncRNA AFAP1-AS1 expression. Further studies verified that PTEN deficiency effectively attenuated the ability of AFAP1-AS1 in promoting ccRCC cell proliferation, invasion, migration, and EMT. Moreover, the similar biological response of silencing AFAP1-AS1 was observed in our ccRCC mice model. Knockdown of AFAP1-AS1 evidently suppressed tumor growth. Taken together, our results provide the evidences that silencing of AFAP1-AS1 inhibits cell proliferation, EMT, and metastasis through PTEN-dependent signaling, and our findings elucidate a novel potential therapeutic target or biomarker for the treatment of ccRCC.     

Cell Kinetics of Head and Neck Squamous Cell Carcinomas: Prognostic implications

Forty-three squamous cell carcinomas of the head and neck region were in vivo labelled with the thymidine analogue iododeoxyuridine. Combined flow cytometric (FCM) and immunohistochemical (IHC) analysis was performed, and the following parameters calculated: labelling index (LI), S-phase time (T_s) and potential tumor doubling time (T_pot). Complete FCM and IHC analyses could successfully be performed in 31 cases, showing a median LI of 13.6% with FCM and 9.1% with IHC. A correlation achieved between LI/FCM and LI/IHC was due to the aneuploid cases, whereas the diploid cases showed no such correlation. Data indicated that T_pot calculated with LI from IHC (T_pot/IHC) might be a prognostic factor, in contrast to T_pot determined using LI/FCM.     

Autocrine and Paracrine Regulation of Neoplastic Cell Growth in Hairy Cell Leukemia

Hairy cell leukemia (HCL), a rare haematological disorder of B-cell origin, mainly presents with bone marrow infiltration, haematopoietic insufficiency, and splenomegaly. In some cases, osteolytic lesions can be observed. Many of these clinical features, especially haematopoietic insufficiency and osteolytic lesions are likely to be caused by soluble factors, such as cytokines. There is evidence that these factors are produced by the malignant hairy cells themselves, suggesting a paracrine pathway. The importance of autocrine as well as paracrine growth loops in growth regulation of HCL-cells is supported by a series of excellent studies, performed within the last few years. It could be clearly shown that cytokines are involved in this autocrine and paracrine regulatory process. The most important cytokines which should be mentioned in this respect are tumor necrosis factor alpha. (TNEa). Interleukin-2 (IL-2), Interleukin-4 (IL-4), Interleukin-6 (IL-6) and B-cell-growth factor (BCGF). The role of other factors such as viruses and oncogenes remains rather unclear. Nevertheless, recent data suggest that the c-fms, which encodes for the macrophage colony stimulating factor (M-CSF) may be involved in the pathophysiological control of HCL growth.     

贝母甲素抑制AML-KG-1a细胞增殖与诱导细胞凋亡研究

[目的]通过体外培养和流式细胞术,检测贝母甲素对人急性髓系白血病细胞KG-1a增殖抑制、诱导凋亡和细胞周期分布的影响。[方法]利用MTT法检测贝母甲素对KG-1a的增殖抑制作用,计算抑制率;以流式细胞术检测贝母甲素对细胞凋亡及周期分布的影响。[结果]1与对照组比较,贝母甲素作用于KG-1a细胞24h、48h、72h的吸光度(OD)值均明显降低(P<0.05),且细胞生长抑制率与药物浓度呈正相关。2与对照组比较,贝母甲素能够提高KG-1a细胞晚期凋亡及总凋亡率,差异均有统计学意义(P=0.003,P=0.001)。3贝母甲素干预KG-1a后,细胞周期发生明显变化,G0/G1期细胞比例降低、G2/M期细胞比例升高、凋亡率升高(P=0.001,P=0.002,P=0.046)。[结论 ]贝母甲素具有抑制具有LSC特性的KG-1a细胞增殖、降低G0/G1期细胞比例、升高G2/M期细胞比例、诱导细胞凋亡效应。