Expression of Polt in tissues and cell lines of transitional cell carcinoma

Objective：To explore the expression of DNA polymerase iota in transitional cell carcinoma ceils and tissues; Methods： RT-PCR was applied to detect the expression of polymerase iota in BIU87 and T24 ceils, then the expression of polymerase iota was also detected in the same way in transitional cell car- cinoma which was derived from clinical bladder carcinoma and renal pelvic carcinoma. Results ： The expres- sion of Poh was low in bladder normal membrana mucosa but significantly elevated in transitional cell car- cinoma ceils. Compared with the expression of polymerase iota in bladder normal mucous membranes, the expression of polymerase iota was significantly increased in transitional cell carcinoma tissue （P〈0. 01 ） and associated with the grade of transitional cell carcinoma. Conclusion： The significantly increased ex- pression of polymerase iota may be associated with the generation and development of transitional cell carcinoma, even with its high heterogenicity.     

Expression of cyclooxygenase-2 mRNA in drug-sensitive cell and drugresistant strains of ovarian cancer cell lines

Objective： To investigate the expression of cyclooxygenase-2 （COX-2） mRNA in drug-sensitive cell and drugresistant clones of ovarian cancer cell lines. Methods： RT-PCR and immunocytochemistry were used to investigate the expression of cyclooxygenase-2 in 3 clones drug-sensitive and 5 clones drug-resistant ovarian cancer cell. Results： Strong COX-2 mRNA expressions were detected in 3 clones of drug-sensitive cell and weak expressions were detected in 5 clones of drug-resistant cell. The protein expression of COX-2 in drug-sensitive cell was strongly positive reaction in immunocytochemistry stain and there was a weak positive reaction in 5 clones of drug-resistant cell. Conclusion： The expression of COX-2 mRNA in drug-sensitive cell strains is much higher than that in drugresistant strains of ovarian cancer cell lines, providing a basis of the chemoprevention for ovarian cancer.     

Establishment and characterization of two cell lines derived from human transitional cell carcinoma.

Two continuous cell lines derived from the neoplastic urothelium had been maintained in culture for more than two years. The first cell line derived from the urothelium of a fusion papillocarcinoma on the left lateral wall of the bladder was designated as TBC-1 and grown in vitro for more than 150 generations. The second cell line derived from the urothelium of a papillocarcinoma in the left renal pelvis was designated as TPC-1 and grown in vitro for more than 100 generations. Characterization studies made on both cell lines showed that the cells had a rapid doubling time, exhibited mul-tilayering and produced tumors in sc of BALB / c. Tumor nodules that produced sc of BALB / c kept similar cellular and pathological features to those of the primary biopsy specimens under light and electron microscopes. TPC-1 cell line exhibited a three-dimensional structure of transitional epithelium on the nylon-mesh disk which was coated with a layer of rat tail collagen. Both TBC-1 and TPC-1 cell lines formed colonies     

Construction and screening of suppression subtractive hybridization library of renal cell carcinoma

Objective To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC). Methods Poly A+ RNA was isolated from RCC lines 786-O(tester) and renal cell(RC) lines HK-2 ( driver), respectiely. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit ( Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced. Results The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly,2 represented unknown genes and the other 48 derived from 36 known genes. Conclusion The quality of the SSH library of human RCC is reliable and is construction is the basis for further screening differentially expressed genes of RCC. 6 refs,4 figs, 1 tab.     

Expression of positive and negative regulators of cell cycle during wound healing

Ｗｏｕｎｄｈｅａｌｉｎｇｉｓｔｈｅｐｒｏｃｅｓｓｂｙｗｈｉｃｈｃｅｌｌｐｒｏｌｉｆｅｒａｔｉｏｎｉｓｓｔｒｉｎｇｅｎｔｌｙｃｏｎｔｒｏｌｌｅｄ ＰｒｏｇｒｅｓｓｉｏｎｔｈｒｏｕｇｈｔｈｅＧ1ｐｈａｓｅａｎｄｉｎｉｔｉａｔｉｏｎｏｆｔｈｅｃｅｌｌｃｙｃｌｅｉｎｍａｍｍａｌｉａｎｃｅｌｌｓｄｅｐｅｎｄｓｏｎｔｈｅｂａｌａｎｃｅｏｆｉｎｔｅｒａｃｔｉｏｎｂｅｔｗｅｅｎｃｅｌｌｃｙｃｌｅｐｏｓｉｔｉｖｅｒｅｇｕｌａｔｏｒｓｓｕｃｈａｓｃｙｃｌｉｎＤ1,ｃｙｃｌｉｎＥ ,ｃｙｃｌｉｎ ｄｅｐｅｎｄｅｎｔｋｉｎａ…     

Restoration of cell polarity and bile excretion function of hepatocytes in sandwich-culture

Objective： To investigate the nature of the restoration of cell polarity and bile excretion function in Sandwich-cultured hepatocytes. Methods ： Freshly isolated hepatocytes from male Sprague-Dawley rats were cultured in a double layer collagen gel Sandwich configuration. Morphological changes were observed under a inverted microscope. The domain specific membrane associated protein DPP IV was tested by immunofluorescence, and the bile excretion function was determined by using fluorescein diacetate. Hepatocytes cultured on a single layer of collagen gel were taken as control. Results.. Adult rat hepatocytes cultured in a double layer collagen gel sandwich configuration regained its morphological and functional polarity and maintained polygonal morphology for at least 4 weeks. Immunofluorescence studies using antibodies against DPP IV showed polarity restoration as early as 48 h. After cultured in the double layer collagen gel Sandwich configuration for 96 h the hepatocytes began to excrete bile; while hepatocytes cultured on a single layer collagen gel had no bile excretion. Conclusion.. Hepatocytes cultured in a double layer collagen gel Sandwich configuration are able to regain their morphological and functional polarity given certain conditions. Hepaotcyte culture is a useful tool for the study of polarity restoration.     

Modulating effects of survivin antisense oligonucleotide on changes of apoptosis and cell cycle of human hepatocellular carcinoma cell line SMMC-7721

Modulating effects of survivin antisense oligonucleotide on changes of apoptosis and cell cycle of human hepatocellular carcinoma cell line SMMC-7721@陈涛$Gener Surg Center,Gener Hosp Chengdu Command,Chengdu 610083     

Expression and significance of PTEN and PCNA in human laryngeal squamous cell carcinoma

TumorsuppressedgenePTEN(phosphotaseandten sionhomologdeletedfromchromsome 10 )wasidentifiedbythreedifferentgroupsin 1997 PTENismutatedinsporad icbrain ,breastandprostatecancer.ThenormalexpressionofPTENcansuppresstumorcellsinvading ,metastasisandgrowth.TheabnormalexpressionofPTENwillresultinPTENproteindecreasingordisappearing .Proliferatingcellnuclearantigen (PCNA )isanauxiliaryproteinofDNApolymerase ,whichisexpressedinG1stageofcellcircle ,getstotheclimaxinSstage ,anddecreasesinG2 -…     

Chemosensitivity of salivary gland and oral cancer cell lines.

Chemosensitivity of salivary gland and oral cancer cell lines to 14 antitumor drugs was tested using a tetrazolium-based colorimetric assay and the relative antitumor activity (RAA) of the drugs was compared. Adriamycin (ADM), methotrexate and fluorouracil (5-FU) showed the most potent RAA against the cell lines while cantharidin did not. The rank orders of other 10 drugs against each cell line differed from each other.

     

Correlation of cytotoxic effect of transmembrane and secretory TNF-α to cell cycle

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G1 phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G1 phase. L929 cells were sensitive to s-TNF-α when synchronized in G1 phase (cytotoxicity 49.8%) while their sensi-tivity to tm-TNF-α was highest in S phase (45.7%) and G1/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.     

Cloning and expression of human arresten gene and effect of its recombinant protein on endothelial cell proliferation

Cloning and expression of human arresten gene and effect of its recombinant protein on endothelial cell proliferation@宋自芳$Dept Gener Surg,Union Hosp,Tongji Med Col,Huazhong Univ Sci Technol,Wuhan 430022     

Experssion of tenascin gene in benign and malignant keratinocyte cell lines and its implication

ＥｘｐｅｒｓｉｏｎｏｆｔｅｎａｓｃｉｎｇｅｎｅｉｎｂｅｎｉｇｎａｎｄｍａｌｉｇｎａｎｔｋｅｒａｔｉｎｏｃｙｔｅｃｅｌｌｉｎｅｓａｎｄｉｔｓｉｍｐｌｉｃａｔｉｏｎＣａｉＺｈｅｎ蔡真ＤｅｐａｒｔｍｅｎｔｏｆＨｅｍａｔｏｌｏｇｙ，ＴｈｅＦｉｒｓｔＡｆｉｌｉ...     

Expression of estrogen receptors and progesterone receptors in transitional cell carcinoma of bladder.

ＥｘｐｒｅｓｉｏｎｏｆｅｓｔｒｏｇｅｎｒｅｃｅｐｔｏｒｓａｎｄｐｒｏｇｅｓｔｅｒｏｎｅｒｅｃｅｐｔｏｒｓｉｎｔｒａｎｓｉｔｉｏｎａｌｃｅｌｃａｒｃｉｎｏｍａｏｆｂｌａｄｄｅｒＳｈａｎＹｕｘｉ单玉喜，ＬｉＬｉｎｇｌｉｎｇ李玲玲，ＹａｎＣｈｕｎｈａｉ颜纯...     

Relationship between microvascular structure and biological characteristics of giant cell tumour of bone.

The vascular architecture of bone giant cell tumor was observed histologically with resin cast technique and scanning elec tron microseopy. Three types of capillaries were noted in the tumor tissue: extruding club-like capillary in the outer zone of the tumor; sinusoid capillary running disorderly in the intermediate zone; cecum capillary in the central zone. The pattern of vascular structure was believed to be correlated with tumor growth.     

Analysis of nuclear DNA content of cells from liver cell dysplasia and hepatocellular carcinoma

Nuclear DNA content of malignant cells was measured by OPTON III microspectrophotometry in 19 cases of hepatocellular carcinoma (HCC), as well as cells from the paraneoplastic liver tissue. The mean DNA value increased greatly from non-liver cells dysplasia (LCD), LCD to HCC (P<0. 05—0. 01), In histograms, non-LCD hepatocytes showed a single peak type located at 2C. HCC showed lower multiple peaks which migrated to 4C and spread widely with more aneuploid (>5C) cells, This histograms of LCD were between non-LCD and HCC,and there were also some aneuploid cells in LCD. These results indicate that LCD is a group of proliferating cells and those with relatively high ratio of aneuploid cells might be premalignant. Recently, many reports on analysis of nuclear DNA content of esophageal, gastric, nasopharyngeal and lung cancer, as well as cells of precancerous lesions have been published. Most results showed that nuclear DNA content of the conditions above mentioned and their number of chromsome were often alterated. Analysis on nuclear DNA content of hepatocellular carcinoma (HCC) and liver cell dysplasia (LCD) are vary rare. And so far, there have been various considerations on behavor of LCD and its relationship to HCC. We would attempt to further approach biologically nature of LCD and its association with HCC by measuring nuclear DNA content.     

Effect of chronic hypoxia on LLC-PK_1 cell proliferatioll and dedifferentiation

Whiletheeffectofacutelschemlaoracutehypoxiaoncellmetabolismandcelldeathhavebeeninvestigatedingreatdetail,littleisknownconcerningthemechanismswherebycellularfunctionsaremodifiedinchronichypoxia.Actually,renovasculardiseases,suchasrenalvasculltis,hypertension,glomerulardiseasesandtubularinterstitialdiseasescancausechronicischemiaandhypoxiaintubularcells.Chronichypoxiaisanimportantfeatureofmanypathophysiologlcconditionsinchronicrenaldiseasesll4j.Toexplorewhatrolechronichypoxiaplaysintileprogressi…     

Genetic variants of miRNA sequences and non-small cell lung cancer survival

Recent evidence indieates that small noneoding RNA molecules known as microRNAs(miRNAs)van function as tumor suppressors and oncogenes.Mutation,misexpression,and altered mature miRNA prueessing are implicated in carcinogenesis and tu- mor progression.Because SNPs in pre-miRNAs could alter miRNA processing,expression,and/or binding to target mRNA,we con- ducted a systematic survey of common pre-miRNA SNPs and their surrounding regions and evaluated in detail the association of 4 of these SNPs with the survival of individuals with non-small cell lung cancer(NSCLC).When we assumed that disease susceptibility was inherited as a recessive phenotype,we found that the rs11614913 SNP in hsa-mir-196a2 was associated with survival in individ- uals with NSCLC.Specifically,survival was significantly decreased in individuals who were homozygous CC at SNP rs11614913.In the genotype-phenoiype correlation analysis of 23 human lung cancer tissue samples,rs11614913 CC was associated with a statisti- cally signifieant increase in mature hsa-mir-196a expression hut not with changes in levels of the precursor,suggesting enhanced processing of the pre-miRNA to its mature form.Furthermore,binding assays revealed that the rs11614913 SNP can affect hinding of mature hsa-mir-196a2-3p to its target mRNA.Therefore,the rs11614913 SNP in hsa-mir-196a2 may be a prognostic biomarker for NSCLC.Further characterization of miRNA SNPs may open new avenues for the study of cancer and therapeutic,interventions.     

Implication of serum levels of interleukin-18 and nitric oxide in tumor growth and metastasis of non-small cell lung cancer

Implication of serum levels of interleukin-18 and nitric oxide in tumor growth and metastasis of non-small cell lung cancer@付向宁$Dept Cardiothorac Surg,Tongji Hosp,Tongji Med Col,Huazhong Univ Sci Technol,Wuhan 430030     

Prevalence of human papillomaviruses 16 and 18 in transitional cell carcinoma of bladder.

It has been speculated that human papillomaviruses (HPV) might be a causative agent of transitional cell carcinoma of the bladder. With the polymerase chain reaction technic, fresh specimens of 53 transitional cell carcinomas of the bladder were examined, and bladder epithelium from 12 patients with hyperplasia of prostate and peripheral blood lymphocytes of 8 normal individuals served as controls. The primers set for HPV-16 and 18 were selected from E6/E7 regions of open reading frames. HPV-16 was positive in 52.8% of the specimens examined, whereas HPV-18 positive in only 3.7%. No controls showed either HPV-16 or 18. Reports on the role of HPVs in carcinogenesis of bladder are scanty and yet there exist conspicuous discrepancies among them. There is a diversity of opinions on the contamination and variation of technical procedures. More work should be done before the relationship between HPV-16 and development of bladder carcinoma can be clarified.

     

Quantitative expression of MMP-2 and FN in high metastatic and low metastatic cell lines of breast cancer

Quantitative expression of MMP-2 and FN in high metastatic and low metastatic cell lines of breast cancer@肖春花$Dept Biochemist Mol Biol,Cancer Hosp,Tianjin Med Univ,Tianjin 300060