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Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix 总被引:48,自引:0,他引:48
A total of eight human epithelial cell lines derived from the carcinoma of the cervix were examined for the presence of human papillomaviruses (HPVs) types 16 and 18 DNA sequences. Six out of eight cell lines contain sequences hybridizing to the DNA of these viruses. Two of the cell lines contain sequences hybridizing specifically to HPV 16. One of these two cell lines contains all of the HPV 16 sequences and the other cell line is missing fragments containing early regions E2 and E4 and some of the late regions. Four of the cell lines contain sequences hybridizing specifically to HPV 18. All these cell lines are missing fragments containing early regions E2, E4, and E5. Interestingly, all the cell lines contain sequences corresponding to early regions E1, E6, and E7. 相似文献
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Myo-Inositol (INO) is a biomolecule with crucial functions in many aspects. In this study, hyperbranched copolymers for gene delivery were synthesized based on inositol and low molecular weight polyethylenimine. The capacity of INO-PEIs to load plasmid DNA and their biocompatibility was demonstrated. A tumor target ligand, folic acid (FA), which was widely used for drug delivery systems, was subsequently conjugated to INO-PEIs and resulted in INO-PEI-FA copolymers. The polymers were then evaluated on their activity to mediate transgene expression in mammalian cell lines. As indicated, INO-PEIs were able to mediate efficient transgene expression, which was particularly noticeable in carcinoma cell line HeLa. INO-PEI-FA further improved the efficiency in HepG2. Distribution of INO-PEI-FA polymers in non-carcinoma NIH 3T3 and carcinoma HeLa cell lines was discussed. 相似文献
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This study quantified the constitutive and interferon-gamma (IFN-gamma) stimulated expression of MHC class I (HLA-ABC and beta 2 microglobulin) and class II antigens (HLA-DR, -DP, -DQ) on normal and malignant oral keratinocytes using radioimmunoassay and immunocytochemical techniques. Normal keratinocytes and three of four malignant cell lines (H103, H157, H314) expressed MHC class I antigens constitutively; IFN-gamma increased MHC class I expression with significant changes in normals, H157 and H314. Normal keratinocytes expressed significantly more constitutive MHC class I antigens than H103 and H157 and significantly more IFN-gamma stimulated MHC class I antigens than H103, H157 and H314. MHC class II antigens predominantly were not expressed constitutively on normals, H103 and H157 but, in H314, HLA-DR, -DP and -DQ antigens were demonstrated on 35, 11 and 5 per cent of cells, respectively, and resulted in a non-coordinated pattern of expression (HLA-DR greater than -DP = -DQ). IFN-gamma induced HLA-DR on normals, H103 and H157, whilst HLA-DP and -DQ remained undetectable. In H314, IFN-gamma enhanced HLA-DR, -DP and -DQ (significant increase of HLA-DQ) but the interrelationship between these antigens was maintained (HLA-DR greater than -DP = -DQ). Normal keratinocytes expressed significantly more IFN-gamma stimulated HLA-DR than H103 and H157 but significantly less HLA-DR than H314 under similar experimental conditions. One oral malignant cell line (H191) did not express MHC class I and MHC class II antigens either constitutively or in response to IFN-gamma. The results demonstrate aberrant patterns of MHC expression (absence, enhanced, diminished) in the different malignant oral keratinocyte cell lines. 相似文献
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The cell lines C-4I and C-4II were established in culture from a nonkeratinizing squamous carcinoma of the uterine cervix. Both lines contain human papillomavirus (HPV) type 18 DNA (Brandt et al., Cold Spring Harbor Laboratories, 5:179, 1987) and both are hypodiploid with similar, but not identical, karyotypes. Each line expresses multiple characteristics of ectocervical epithelial differentiation, but the characteristics differ between the lines. In the present study, G banding of the lines showed that cells of both lines have two normal chromosomes 1-5, 8-10, 13, 16, and 17, one normal chromosome 12 and 14, and no normal chromosomes 15 and 18. The lines share three abnormal chromosomes, der(8)t(8q;12q), der(18)t(18q;?), and i(5p). There are specific differences between the lines. C-4I has two normal chromosomes 6, while C-4II has one; C-4II has two chromosomes 11 and der(18)t(18q;?), while C-4I lacks both chromosomes 11 and has one der(18)t(18q;?). Each line has unique markers that include del(11)(p11), del(22)(q12), and del(21)(q21) in C-4I and i(15q), der(X)t(Xq;9p), der(6)t(6p;14q), and del(4)(q21) in C-4II. The results show that these phenotypically distinct lines are derived from the same clone and that the 8q arm (the site of HPV 18 integration) is present in three copies in both lines. They also define several chromosome rearrangements that are compatible with the expression of specific differentiation markers. 相似文献
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IFN-α对宫颈癌细胞MHC-I类链相关蛋白A表达的上调作用 总被引:2,自引:0,他引:2
目的:观察IFNα对宫颈癌细胞MHCI类链相关蛋白A(MICA)表达及功能的影响。方法:以IFNα诱导宫颈癌细胞系HeLa和Caski后,用半定量RTPCR和免疫组化染色法观察诱导前后MICAmRNA和其蛋白表达的变化,用MTT比色法检测诱导前后宫颈癌细胞对外周血NK细胞杀伤敏感性的变化。结果:经IFNα诱导后,HeLa和Caski细胞中MICAmRNA和其表面MICA蛋白的表达水平均明显上调,并呈一定的时间和剂量依赖关系。NK细胞对MICA表达较高的HeLa细胞的杀伤活性,明显高于对MICA弱表达的Caski细胞。经1×106U/LIFNα诱导3d后,两种细胞对NK细胞杀伤的敏感性均明显增强(P<0.01)。结论:IFNα可上调MICA在肿瘤细胞表面的表达,并可增强肿瘤细胞对NK细胞杀伤的敏感性。 相似文献
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Jia-feng Niu Cai Zhang Jian-hua Zhang Xiao-qun Xu Wei-dong Zhang Jun-fu Wang Chun-yi Gao 《细胞与分子免疫学杂志》2006,22(5):557-559
AIM: To investigate the effect of IFN-alpha on expression of MHC class I chain-related protein A (MICA) in the cervical carcinoma cell lines. METHODS: The cervical carcinoma cell lines (HeLa and Caski) were treated with IFN-alpha. The expression of MICA was measured by RT-PCR and by immunohistochemical staining. The cytotoxicity of human NK cells to the IFN-alpha treated cervical carcinoma cells was detected by MTT method. RESULTS: The mRNA and protein expression of MICA was up-regulated by IFN-alpha in HeLa and Caski cells in a time and dose-dependent manner. Compared to Caski cells which weakly expressed MICA, higher cytolytic activity of NK cells was found against HeLa cells, which expressed relatively higher level of MICA. After being treated with IFN-alpha for 3 d, the susceptibility of the two cervical carcinoma cells to NK cytolysis was increased significantly. CONCLUSION: IFN-alpha can up-regulate the MICA expression in the cervical carcinoma cell lines and thereby enhance the susceptibility to cytolysis of NK cells. 相似文献
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Repression of human papillomavirus (HPV) E6 and E7 oncogenes in established cervical carcinoma cell lines causes senescence due to reactivation of cellular tumor suppressor pathways. Here, we determined whether ongoing expression of HPV16 or HPV18 oncogenes is required for the proliferation of primary human cervical carcinoma cells in serum-free conditions at low passage number after isolation from patients. We used an SV40 viral vector expressing the bovine papillomavirus E2 protein to repress E6 and E7 in these cells. To enable efficient SV40 infection and E2 gene delivery, we first incubated the primary cervical cancer cells with the ganglioside GM1, a cell-surface receptor for SV40 that is limiting in these cells. Repression of HPV in primary cervical carcinoma cells caused them to undergo senescence, but the E2 protein had little effect on HPV-negative primary cells. These data suggest that E6 and E7 dependence is an inherent property of human cervical cancer cells. 相似文献
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The distribution of 1,912 breakpoints observed in a series of 148 cervical cancers was analyzed. Fifty bands were shown to be nonrandomly involved in chromosome structural rearrangements. One hundred thirty-three breaks were noted in bands known to contain a human papillomavirus integration site, and 454 breaks were noted in bands containing a herpes simplex virus breakage site. We suggest that herpes simplex viruses and, possibly, papillomaviruses play an important role in the carcinogenesis and/or development of cytogenetic abnormalities in cervical cancers. 相似文献
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Detection of human papillomavirus DNA sequences by in situ DNA-DNA hybridisation in cervical intraepithelial neoplasia and invasive carcinoma: a retrospective study. 总被引:1,自引:3,他引:1
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Human papillomavirus (HPV) infection of cervical intraepithelial neoplasia (CIN) and invasive cervical carcinoma was investigated using in situ DNA-DNA hybridisation on histological sections of formalin fixed, paraffin embedded tissue to assess the technique's sensitivity and to assess retrospectively the association between HPV16 and invasive cervical carcinoma. HPV DNA was detected in 16 of 33 biopsy specimens of CIN. Cells containing viral DNA were more numerous than those positive for viral structural proteins. HPV DNA was also present in less differentiated cells deeper in the epithelium. The detection rate in CIN was lower than that reported for other hybridisation techniques such as Southern blotting. In a retrospective study of biopsy specimens of invasive squamous carcinoma of the cervix HPV16 DNA, the virus most commonly associated with cervical malignant disease, was found in 20 of 25 cases, including those dating from as far back as 1932. The level of sensitivity was similar to that reported for other hybridisation techniques. DNA positive cells were focally distributed in the invasive tumours, and most tumour cells were negative for viral DNA, a result consistent with the low copy number found in malignant cells. It is concluded that HPV16 is not a new virus but that its prevalence is a result of changes in sexual behaviour and that in situ hybridisation is useful in the localisation of HPV DNA replication in CIN and invasive carcinoma. 相似文献
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目的探讨宫颈上皮内瘤变(CIN)中不同类型人乳头瘤病毒(HPV)的感染及分型情况。方法回顾性研究北京军区总医院妇科门诊134例已经确诊的CIN患者,采用凯普的核酸分子快速导流杂交基因芯片技术对HPV进行分型。结果 1.各级别CIN以HPV高危亚型单一或者多重感染为主;2.在HPV阳性的101例患者中,单一感染者73例(72.28%),双重感染者24例(23.76%),三重感染者3例(2.97%),四重感染者1例(0.99%);3.HPV-16、58感染率最高,分别为30.83%,24.06%。结论 HPV感染以高危型和多重感染为主;随CIN级别的增高,HPV阳性率也是增高的;HPV-16亚型的感染率最高。 相似文献
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A. A. WADEE A. PATERSON K. A. COPLAN S. G. REDDY 《Clinical and experimental immunology》1994,97(2):328-333
The present study undertook to investigate the biological significance of human leucocyte antigen expression in hepatocellular carcinoma and to elucidate the role of potential modulating agents on human leucocyte antigen expression. These studies used several hepatic tumour-derived cell lines as in vitro model systems. The cell lines included PLC/PRF/5 (Alexander cell line), Hep3B. HepG2, TONG PHC, HA22T/VGH, HA59T/VGH and Mahlavu, The cell lines K562 and Raji were used as negative and positive controls, respectively. K562, a B tymphoid-derived cell line, was shown to express negligible amounts of human leucocyte anligens, while Raji, an erythromyeloid-derived cell line, expressed both class I and class II human leucocyte anligens as well as their respective invariant chains, β2-niicroglobulin and Ii. Using an ELISA, experiments performed on these cell lines confirmed the natural expression of class I and class II antigens by the HA22T/VGH and HA59T/VGH cell lines, whereas PLC/PRF/5 displayed class II surface antigens only. The effects of modulating agents such as interferon-gamma sodium bulyrate and clofazimine on human leucocyte antigen expression were investigated using the HA22T/VGH, HA59T/VGH and TONG PHC cell lines. These agents increased class 1 and class II human leucocyte antigen expression on HA22T/VGH and TONG PHC cells, but had no effect on the HA59T/VGH cell line. The results suggest a potential use for these agents as modulators of human leucocyte antigen expression by human heptocellular cell lines. 相似文献
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Chromosomal integration sites of human papillomavirus DNA in three cervical cancer cell lines mapped by in situ hybridization 总被引:52,自引:0,他引:52
Metaphase chromosomes of three cervical cancer cell lines (HeLa, CasKi, SiHa) were subjected to in situ hybridizations with the DNA of human papillomaviruses (HPV) types 16 and 18, respectively. Previous studies have demonstrated multiple copies of HPV 18 DNA in HeLa and of HPV 16 DNA in CasKi cells, but only 1–2 HPV 16 copies in cells of the SiHa line. The viral DNA persists in an integrated state (Schwarz et al 1985). Analysis of the integration sites revealed at least 11 chromosomal sites of HPV 16 integration in CasKi cells. SiHa cells contain integrated HPV 16 DNA in the region q21–q31 of chromosome No. 13. In HeLa cells integration of HPV 18 occurred in chromosome No. 8, band q24. Thus, no evidence was obtained for the existence of preferential chromosomal regions for HPV integration. The data indirectly support a trans-acting function of HPV-mediated cell transformation.On leave of absence from the Institute of Infectious and Parasitic Diseases, Medical Academy, VI. Zaimov 26, 1504 Sofia, Bulgaria 相似文献