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Biomedical subjects

Feng Wang-Johanning

Publications and source records attributed to Feng Wang-Johanning.

5 recordsLinked to original sources

Expression of multiple human endogenous retrovirus surface envelope proteins in ovarian cancer.

Individual classes of human endogenous retrovirus (HERV) genes and proteins are expressed in cancer, but expression of more than one type of HERV is rare. We report here the expression of multiple HERV genes and proteins in ovarian cell lines and tissues. Expression of HERV-K env mRNA was greater in ovarian epithelial tumors than in normal ovarian tissues (N = 254). The expression of this protein on the surface and in the cytoplasm of ovarian cancer cells was confirmed using anti-HERV-K specific antibody by flow cytometric analysis. The frequency of expression of HERV-K env protein in multitissue microarrays (N = 641) was determined by immunohistochemistry and a significant correlation with tumor histotype was found. A significantly increased expression of HERV-K was observed in tumors with low malignant potential and low grade, relative to expression in normal ovarian tissues. The increase in expression of HERV-K env protein took place in a stepwise fashion in serous papillary adenocarcinoma. Interestingly, we found that other classes of HERV env mRNAs, including ERV3 and HERV-E, are expressed in the same ovarian cancer tissues that expressed HERV-K. Furthermore, anti-HERV antibodies including anti-ERV3 (30%), anti-HERV-E (40%) and anti-HERV-K (55%) were detected in patients with ovarian cancer, but not in normal female controls. HERV env proteins are frequently transcribed and translated in ovarian epithelial tumors, and multiple HERV families are detectable in ovarian cancer. HERV env proteins, and especially those expressed on the cell surface, may serve as novel tumor targets for detection, diagnosis and immunotherapy of ovarian cancer.

Adenocarcinoma, Clear Cell↗

Viral detection.

The focus of this chapter is the detection of DNA viruses. The emphasis is on amplification reactions that include reverse transcription-polymerase chain reaction (RT-PCR), PCR, real-time RT-PCR, and real-time PCR methods. Amplification of the E6 and E7 oncoproteins of HPV16 is described in detail, and primers and probes that can be used to amplify these oncogenes are described. Techniques to quantify these oncogenes in infected human tissue specimens are presented, and analysis of data resulting from real-time PCR detection of the E6 and E7 oncogenes is discussed. Other methods for viral nucleic acid detection, including nested PCR amplification, ligase chain reaction, and enzyme-linked immunosorbent assay (ELISA), are also briefly discussed.

DNA Viruses↗

Detecting the expression of human endogenous retrovirus E envelope transcripts in human prostate adenocarcinoma.

BACKGROUND: The expression of human endogenous retrovirus (HERV) mRNA and proteins was associated recently with diseases that include human malignancies. The authors report that, in the current study, transcripts encoding the envelope region of an HERV family, HERV-E, were expressed in human prostate carcinoma. METHODS: RNA was isolated from various prostate tissues and was tested for the expression of various HERV envelope (env) genes by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, RNA in situ hybridization (ISH), and Northern blot analysis. Variants of HERV that appeared in prostate carcinoma tissues were sequenced, and HERV-E was expressed in prokaryotic and eukaryotic systems. RESULTS: In the current study, the authors found that the mRNA of the env gene of one particular family of HERVs, HERV-E, was expressed in some prostate carcinoma tissues (38.8% positive; n = 49 specimens) but not in normal prostate tissues using RT-PCR, RNA ISH, and Northern blot assays. The expression of HERV-E transcripts in prostate tumor epithelial cells was confirmed further by ISH using an HERV-E specific antisense probe. Approximately 50% of the cDNA of HERV-E obtained from prostate carcinoma specimens contained no stop codon and expressed proteins in prokaryotic or eukaryotic expression systems. Furthermore, the expression of both HERV-E and ERV3 (another class of HERV) was detected in the same prostate carcinoma tissues. CONCLUSIONS: The expression and distribution of multiple HERV-E endogenous retroviral elements in prostate carcinoma, but not in normal control specimens, suggests that they may serve as novel tumor markers for the early diagnosis and immunotherapy of patients with prostate carcinoma.

Blotting, Northern↗

Quantitation of HERV-K env gene expression and splicing in human breast cancer.

Human endogenous retroviruses (HERVs) comprise up to 8% of the human genome. In previous studies, we demonstrated that type 1 HERV-K envelope (env) transcripts are expressed in most human breast cancers, but not in normal breast tissues. In the current study, we report that type 2 HERV-K env transcripts are also present in human breast cancers. By real-time RT-PCR, the expression of HERV-K env transcripts was 5-10-fold higher in breast cancer cell lines treated with estradiol and progesterone than in cells without treatment, and expression was significantly higher in most breast cancer tissues than in normal breast tissues. Furthermore, both types of HERV-K env transcripts were capable of being spliced into subgenomic env transcripts and various splice donor and acceptor sites were detected in breast cancers. The selective expression and distribution of multiple HERV-K endogenous retroviral element splice variants in breast cancer, but not in normal controls, suggests that they are novel breast tumor markers.

Breast Neoplasms↗

Quantitation of human papillomavirus 16 E6 and E7 DNA and RNA in residual material from ThinPrep Papanicolaou tests using real-time polymerase chain reaction analysis.

BACKGROUND: The detection of specific human papillomavirus 16 (HPV-16) E6 and E7 oncogene transcripts may be a sensitive indicator of the direct involvement of viral oncogenes in the development of cervical neoplasia and carcinoma. The goal of this study was to determine the potential clinical uses of real-time polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR) methods for evaluating HPV-16 E6 and E7 oncogene expression. METHODS: ThinPrep cervical samples were tested for expression of oncogenes of HPV-16 by real-time PCR or RT-PCR analysis and were compared with detection of expression by conventional PCR and RT-PCR analysis. Both sets of results were correlated with the cytologic diagnosis of the cervical samples. RESULTS: The presence of HPV-16 E6 and E7 DNA and RNA was observed only in HPV-16 positive cervical carcinoma cell lines but not in HPV-18 positive or HPV negative cell lines. The percentage positive for HPV-16 E6 or E7 DNA in a series of ThinPrep cervical cytologic samples (n = 348 samples) was 0% for negative samples (n = 45 samples), 9.7% for atypical squamous cells of undetermined significance (ASCUS; n = 144 samples), 16.9% for low-grade squamous intraepithelial lesion (LSIL; n = 118 samples), and 51.2% for high-grade intraepithelial lesion (HSIL; n = 41 samples). The copy numbers per nanogram for both DNA and RNA E6 and E7 were increased significantly as severity of the lesions progressed from ASCUS to HSIL, and RNA copy numbers were a more sensitive indicator of HPV-16 E6 and E7 expression than DNA copy numbers. The increase in copy numbers took place in a stepwise fashion from ASCUS, to LSIL, to HSIL. CONCLUSIONS: The detection of HPV-16 E6 and E7 expression by real-time RT-PCR or PCR analysis in ThinPrep cervical cytologic specimens may serve as a quick, reliable, and sensitive tool to identify a subset of patients who express HPV-16 oncoproteins.

Adolescent↗