IL-1α and ATP mRNA expression after ultraviolet irradiation in human keratinocyte original-SCC 12F cells

Background Generally speaking, undifferentiated keratinocytes may synthesize a larger amount of IL-α and its production is decreased as cells complete differentiation gradually. Ultraviolet B (UVB)light can signigicantly stimulate the production and release of some cytokines. In this study we investigated the influence of UVB irradiation on IL-1α and adenosine triphosphoric (ATP) mRNA expressions in the human keratinocyte (KC) of original squamous cell carcinoma line (SCC 12F cells).Methods The cultured SCC 12F cells were irradiated with 30 mJ/cm^2 of UVB. Northern blot was employed to analyze the expression of IL-1α and ATP mRNA.Results There was a constitutive expression of IL-1α mRNA in SCC 12F cells. The expression increased in culturing time in regular KC medium and reached the highest expression at 120 hours.The expression level of IL-1α was up-regulated with two peaks at 6 hours and 72 hours, respectively after UVB irradiation. In comparison with IL-1α mRNA expression, ATP mRNA was down-regulated,with similar biphasic peaks, compared with the sham irradiated group.Conclusions SCC12F cells may express IL-1α mRNA constitutively. After UVB irradiation, the mRNA expression of IL-1α and ATP will show opposite effect because of inflammation/immunity and energy consumption mechanisms.     

Expression and characterization of Mac-1-FP fusion protein in CHO cells

To construct mammalian cell expression vectors for Mac-1 with CFP and YFP and apply FRET to study the dimerization and function of CD1 lb( Mac-1 α subunit) and CD18( Mac-1 β subunit). Methods: The mammalian cell expression vector for CD1 lb fused with CFP at the carboxyl terminal was constructed to create recombinant plasmid of pCD11b-CFP. Then pCD11b-CFP was co-transfeeted with pYFP-CD18 into CHO cell, a fibroblast like cell line, as a target cell within which there are some signal pathways involved in inflammatory stimulation but without endogenous Mac-1. Then CHO cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were selected by Western blot and laser scanning confocal microscope. Results: The cyan and yellow fluorescence in co-transfected positive CHO cells were observed under a fluorescence microscope. CHO-Mac-1-FP cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were obtained as demonstrated by Western blot successfully. The adhesive activity of CHO-Mac-1-FP cells with CHO-ICAM-1cells was increased markedly bv treatment with PMA, suggesting the translocation of CD11b-CFP and YFP-CD18 to the plasma membrane in CHO-Mac-1-FP cells and dimerization of CD11b-CFP and YFP-CD18 just as the function of the wild type Mac-1. Conclusion: CHO-Mac-1-FP cells with adhesive activity are established successfully, thus CHO-Mac-1-FP cells may be useful for the study of Mac-1 by FRET and for other purposes.     

Expression and significance of 34βE12 and P63 in breast cancer

目的 研究高分子细胞角蛋白(34βE12)及肌上皮标记物(P63)在乳腺良恶性组织中的诊断意义.方法 用免疫组织化学方法(Envision)检测34βE12及P63在46例乳腺增生症组织和86例乳腺癌组织中的表达水平.结果 34βE12在乳腺良性增生症组织中阳性表达率100％,在乳腺癌组织中阳性表达率38.4％,34βE12的表达率及其强度在良性增生症与癌组间比较差异有统计学意义(P＜0.05).46例乳腺增生症组织P63表达均为阳性,86例浸润性癌,P63均为阴性.结论 通过对乳腺病变的观察,腺体周围基底细胞是否存在是乳腺良恶性的重要鉴别点,34βE12和P63是基底细胞特异性标记物,而P63更敏感,是诊断乳腺癌的首选抗体,而两者同时选用则更为理想.     

Effects of TNF-α and curcumin on the expression of VEGF in Raji and U937 cells and on angiogenesis in ECV304 cells

Background To better understand the possibilities of antiangiogenic tumor therapy and to assess possible side effects, we investigated the effect of tumour necrosis factor （TNF）-α and curcumin on the expression of vascular endothelial growth factor （VEGF） in U937 and Raji cell lines and their effect on angiogenesis in a human umbilical vein endothelial cell （HUVECs）-derived cell line （ECV304）, and also the relationship between Notchl and VEGF. The aim of this study was to elucidate potential mechanisms controlling tumor neovascularization. Methods VEGF secreted by U937 and Raji cell lines was determined by ELISA. Angiogenesis was tested by network formation of endothelial cells on Matrigel. Levels of VEGF mRNA in U937 and Raji cells and Notchl mRNA levels in EV304 cells were determined by RT-PCR. Results Secretion of VEGF by U937 and Raji cells was increased by TNF-α treatment and suppressed by curcumin （P 〈 0. 01 ）. The mRNA expression of VEGF165 and VEGF121 （containing 165 and 121 amino acid residues, respectively） were detected in any fractions. TNF-α augmented the expression of VEGF165 and VEGF121 mRNA and curcumin reduced the expression （P 〈0. 01 ）. No networks or cords formed in control and curcumin groups. There was tube formation on matrigel in the supernatants of the Raji culture group and the supernatants groups treated by VEGF group and TNF-α in Raji cell. Notch1 mRNA was detected but there was no significant change in the VEGF group compared with control （P 〉 0. 05）. Conclusions Expressions of VEGF mRNA in U937 and Raji cells were increased by TNF-α and suppressed by curcumin. VEGF and TNF-α can induce angiogenesis, and curcumin can inhibit angiogenesis in ECV304 cells.     

Expression of p63 and Cyclooxygenase-2 and Their Correlation in Skin Tumors

To study the expression of p63 and cyclooxygenase-2 (cox-2) in skin tumors and evalu- ate the correlation between p63 and cox-2, the expressions of cox-2 and p63 were measured by streptavidin-peroxidase complex immunohistochemical technique in 17 cases of skin squamous cell carcinoma(SCC), 19 cases of Bowen's disease(Bowen), 11 cases of actinic keratosis(AK), 12 cases of seborreic keratosis(SK) and 13 specimens of normal skin. Our results showed that the expression of p63 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while the expression of p63 in seborreic keratosis was sig- nificantly higher than that in normal skin. The expression of cox-2 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while no statistical difference was noted in the expression of cox-2 between seborreic keratosis and normal skin. Cox-2 expression was positively correlated with the high p63 expression in malignant skin tu- mors. The increased expression of cox-2 and p63 may play an important role in the development of skin tumors and work synergetically in malignant skin tumors.     

Overexpression of amyloid precursor protein inhibits neurite outgrowth and disrupts cytoskeleton in N2a cells

There is considerable evidence suggesting that altered metabolism of β-amyloid precursor protein (APP) and accumulation of its β-amyloid (Aβ) fragment are key features of Alzheimer‘ s disease (AD). APP is a type Ⅰ integral membrane protein and consists of 695 - 770 amino acids encoded by differentially spliced mRNAs transcribed from a single gene located on human chromosome 21. The 695-amino acid APP is expressed preferentially in the brain. Aβ, the major component of senile plaques, is derived by proteolytic processing of APP by β-and γ-secretase and is constitutively released from most cells.     

The expression of TRAIL and its receptors in osteosarcoma cells and the apoptosis effect of a combination of TRAIL, adriamycin and IFN-γ on MG-63 cells

Objective: This study investigated the effects of rh-TRAIL with or without chemotherapeutic drugs on the apoptosis of the osteosa-rcoma cell line, MG-63, and the influence of chemotherapeutic drugs on changes in the expression of DR5 and YinYang1(YY1) in MG-63 cells. Methods: The effects of treatment with rh-TRAIL alone and/or chemotherapeutic drugs on MG-63 cell growth inhibition and apoptosis were measured using the MTT assay, FACS analysis of Annexin V labeled cells, and the mRNA changes of DR5 and YY1 were detected by RT-PCR. Results: Rh-TRAIL protein inhibited the growth of MG-63 cells, and this inhibition was increased by adriamycin and IFN-γ. Adriamycin and IFN-γ significantly facilitated the induction of the expression of DR5 and reduced the expression of YY1. Conclusion: The apoptosis-inducing effect of rh-TRAIL in MG-63 cells was enhanced by chemotherapeutic drugs.     

The determination and significance of C/EBP in cells of liver cancer and different liver tissues

C/EBP is a sequence-specific DNA-binding protein.In order to identify its distribu-tion,localization and function,immunocytochemical technique(ABC method)was done usinganti-C/EBP polypeptide antibodies 1103~#,425~# in liver specimens from 20 normal adult,5neonatal,6 patients with hepatitis,25 patients with liver cirrhosis,80 patients with hepatocellu-lar carcinoma(40 cases were associated with surrounding nontumorous tissues)and 26 patientswith cholangiocarcinoma(15 cases were associated with surrounding nontumorous tissues).Theresults showed that C/EBP was diffusely distributed in nuclei and cytoplasm of differentiated liv-er cells and very low or undetectable in liver cancer cells.The expression of C/EBP was in pro-portion to differentiated degree of tumor cells,and was obviously weaker than that in surround-ing nontumorous tissues.C/EBP positive staining has also been found in regenerating epithelialcells of bile ductules.The results suggested that C/EBP might play an important role in estab-lishing and maintaining the differentiation of liver cells and might exert an inhibiting effect againsttransformation of liver cells and proliferation of neoplastic tissue.     

The expression of P63 protein in some keratinocyte original tissues and cells

ObjectiveTo examine the expression patterns of p63 in tissues of particular keratinocyte original hyperproliferate diseases and variety cell types for determining if P63 is the marker of proliferative potential keratinocytes.MethodsP63 protein was detected and analyzed by immunoreactivity method and Western blot in biopsy specimens of keratinocyte original disorders including squamous cell carcinomas SCC, basal cell carcinomas BCC, Bowen's disease and other tissues or cells, such as psoriasis vulgaris, normal skin tissues, primary cultured keratinocytes, immortal HaCaT cells, and epidermoid carcinoma cells A431.ResultsP63 protein was expressed in the nuclei of basal and suprabasal layer of the epidermis, germinative cells of sebaceous glands in normal epidermal. P63 was strongly and diffusely detected in the majority of tumor cells in BCC and poorly-differentiated SCC. In Bowen's disease, p63 expresses are remarkable in all cell layers. In the psoriasis plaque epidermal, p63 expressed mainly in basal cells and part of spinous cells. P63 expressed more strongly in primary cultured keratinocytes than in A431 cells or HaCaT cells.ConclusionP63 is a nuclei marker of undifferentiated keratinocytes with the proliferative potential and may disrupt the terminal differentiation. The overexpression of p63 reflects immaturity of the tumor cells. The immunohistochemical staining of p63 may be useful for investigating the origin and differentiation of tumor cells.     

胃癌组织P53、P63蛋白表达及细胞凋亡的研究

目的：探讨P53、P63蛋白表达及细胞凋亡在胃癌发生发展中的可能机制。方法：选择57例手术切除的胃癌及胃癌旁组织,用免疫组化方法检测组织中的P53、P63蛋白表达,同时用Tunel方法检测胃癌组织中的凋亡细胞。结果：胃癌组织P53、P63蛋白表达率显著高于胃癌旁组织（P〈0.05）;低、未分化型腺癌表达率显著高于高、中分化型腺癌（P〈0.05）;有淋巴转移显著高于无转移组（P〈0.05）;P53、P63高表达的癌组织,细胞凋亡指数减少,早期胃癌低于晚期胃癌（P〈0.05）。结论：胃癌组织P53、P63蛋白高表达,可能延缓细胞凋亡;细胞凋亡机制障碍可能是胃癌增生失控的基础。     

Bim蛋白在原发性肝细胞癌中的表达及临床意义

目的：探讨促凋亡蛋白Bim基因在肝癌组织中的表达及其与肝癌临床病理、预后的相关性。方法：收集南通大学附属医院病理科80例原发性肝细胞癌及癌旁组织标本,应用免疫组化Envision法检测Bim蛋白的表达,结合临床病理参数和随访资料进行统计学分析。结果：免疫组化结果显示,在肝癌组织中Bim表达阳性率为15%（12/80）,明显低于癌旁正常肝组织的83.75%（67/80）,两者之间比较差异有统计学意义（χ2=13.714,P<0.001）。Bim蛋白在原发性肝细胞癌中的表达与TNM分期有关（χ2=7.521,P=0.023）。Kaplan-Meier生存曲线显示Bim低表达组比高表达组5年生存率低。结论：Bim蛋白的表达与人原发性肝细胞癌的发生、发展存在一定的关系,可能成为肝细胞癌潜在的治疗靶点。     

丙型肝炎病毒核心蛋白抑制肝癌细胞P16、P27的表达

目的：探讨HCV核心蛋白对肝瘤细胞P16、P27表达的影响。方法：将HCV核心基因cDNA插入真核表达载体PBK－CMV的Hind　Ⅲ和BamH I位点间，构建重组质粒PBK－HCVc。再将重组质粒PBK－HCVc和空载体分别导入肝癌细胞株HepG2中，G418筛选，RT－PCR、蛋白印迹鉴定HCV核心蛋白表达。免疫组化，蛋白印迹检测P16、P27蛋白表达；RT－PCR检测P16、P27mRNA。结果：重组质粒PBK－HCVc在HepG2细胞中有稳定表达；表达HCV核心蛋白的细胞HepG2-C的P16、P27在蛋白水平和mRNA水平较转染空载体的细胞HepG2-CMV明显下降。结论：HCV核心蛋白能抑制P16、P27表达，可能与肝细胞的癌变有关。     

抑癌基因P16在膀胱移行细胞癌中的表达及临床意义

目的　探讨抑癌基因P16蛋白在膀胱移行细胞癌中的表达。方法　采用S -P放射免疫法检测 3 2例膀胱移行细胞癌、10例正常膀胱粘膜中P16蛋白的表达。结果　 3 2例膀胱移行细胞癌中 ,P16蛋白表达总阳性率为 40 .63 % ;在膀胱移行细胞癌Ⅰ、Ⅱ、Ⅲ级中分别为 63 .64 %、3 3 .3 3 %、2 5 .0 0 %。P16蛋白表达的阳性率随病理分级增高而降低 ,Ⅰ级与Ⅱ级、Ⅲ级比较差异无显著性 (P >0 .0 5 )。结论　P16蛋白检测有助于对膀胱移行细胞癌的诊断 ;P16蛋白的缺失与膀胱移行细胞癌的发生、发展和预后有关。     

乳腺癌中Survivin蛋白的表达及其与P53的关系

目的观察Survivin在正常乳腺组织、一般增生、不典型增生及乳腺癌组织中的表达情况,探讨其与突变型P53基因表达的相关性.方法用免疫组化法检测36例乳腺癌组织、30例乳腺增生组织及7例正常乳腺组织中的Survivin和P53的表达.结果在正常乳腺组织→一般增生→不典型增生→乳腺癌的发展过程中,Survivin及P53阳性表达率随乳腺组织增生程度的加重而升高,两者的表达与乳腺癌的组织类型、临床分期和淋巴结转移状况无显著相关.Survivin在P53阳性的乳腺癌组织中的表达率为100%,明显高于P53阴性组.结论Survivin蛋白的异常表达与乳腺癌的发生与发展密切相关,Survivin蛋白表达与P53基因突变显著相关,两者可能通过某种作用共同参与乳腺癌的发生与发展.     

应用组织芯片检测Bmi-1及P16在胆管细胞癌中的表达及意义

目的:探讨Bmi-1及P16基因表达与胆管癌发生、发展的关系。方法:本文采用免疫组织化学技术SP法检测人胆管细胞癌组织芯片150芯(其中包括70例胆管细胞癌组织和癌旁组织5例)标本的Bmi-1及P16蛋白的表达。结果:在胆管细胞癌组织及癌旁组织中,Bmi-1和P16蛋白显著差异表达。结论:Bmi-1在胆管细胞癌组织中呈高表达,而P16蛋白在胆管细胞癌组织中呈低表达,且Bmi-1与P16蛋白的表达呈负相关,可能与胆管癌的发生、发展有关,与预后是否有关,尚需进一步的研究。