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O.151 DNA-image cytometry improves assessment of resection margins
S38
Journal of Cranio-Maxillofacial Surgery 36(2008) Suppl. 1
Tuesday, 9 September 2008, 16.00–17.00
Sala Italia
Cancer biology and epidemiology I O.147 cAMP-phosphodiesterase signal in malignant melanoma cells Y. Watanabe, T. Murata, K. Shimizu, K. Mizoi, J. Nomura, T. Tagawa. Department of Oral and Maxillofacial Surgery, Division of Reparative and Regenerative Medicine, Institute of Medical Science, Mie University Graduate School of Medicine 1, Tsu, Japan Objectives: Cyclic AMP (cAMP) acts as a second messenger in cells and modulates many physiological processes. Intracellular cAMP content is regulated through synthesis by adenylate cyclase and degradation by phosphodiesterases (PDEs). However, no data are available as to the cAMP-PDE signal in malignant melanoma cells. In this study, we examined cAMP-PDE signal in malignant melanoma cell line. Methods: Cell growth: Mouse B16 melanoma cells were plated at 100 cells/well in a 96-well plate and allowed to adhere for 24 h. The cells were cultured in the absence or presence of different concentration of cAMP analog, 8-bromo cAMP, or cGMP analog, 8-bromo cGMP, for 6 days. The cell numbers were calculated using CellTiter 96® Aqueous One Solution Cell Proliferation Assay (Promega, Madison, WI). ˚ cells/25oz. flask. PDE activity: B16 cells were seeded at 1×10A After 3 days, the cells were incubated with 8-bromo cAMP for 15 min, harvested and homogenized in 1 ml of ice-cold homogenization buffer. PDE activity was measured. Results: 8-bromo cAMP inhibited the growth of B16 cells, but not 8-bromo cGMP. And PDE activity in B16 cells was stimulated by 8-bromo cAMP. Conclusions: These data suggested that cAMP-PDE singal was related with the growth of malignant melanoma cells and cAMPPDE singal might be target for an antitumor drug of malignant melanoma. O.148 Chemical surgery for oral cancer S. Kim. Sacred Heart Hospital, Anyang, Republic of Korea Objective: The control of microinvasion is highly important to improve complete cure rate in oral cancer. We wanted to evaluate the therapeutic effects of chemical agent that has sharp threshold in inducing apoptosis of cancer. Methods: Eight patients were included in this clinical trials and six patients might have remained cancer in resection margin. Topical 4-HR application was done for 1 month postoperatively. Results: Seven patients showed tumor free and mean follow-up was 32 months. There was no metastatic tumor after operation. Conclusion: Though this was a preliminary trials, the chemical surgery could be considered as an additive technique to improve cure rate of oral cancer. O.149 Choice of operation volume in glossal cancer T1-3NxMO R. Bakirov, S. Osokin, R. Osokina. Bashkir Clinical Oncology Health Center, UFA, Russia The aim of our research is to evaluate effectiveness of lymph node dissection as an approach which allows giving prognosis of metastasizing in remote postoperative period. We analyzed history cases of 256 patients with glossal cancer T1−3NxMo who received surgical treatment in Head and Neck Tumor Department. We found out that patients who received treatment only within gemiglossectomy had regional metastases in jugular lymph nodes in 28% of cases.
Abstracts, EACMFS XIX Congress The second group of patients is those who received treatment of initial lesion having simultaneous removing of regional lymph drain ducts whose metastasizing process wasn’t proved by the results of diagnostic study. In this group of patients the frequency of metastases to remote lymph nodes was 23%. In 88.4% of operated patients of this group there were no metastases within one year. The patients of the third group in 10.5% of cases had metastases within one year after operation. Simultaneous regional lymph node dissection and operation of the initial lesion T1−3NxMo can be treated as a factor of surgical prognosis of further course of disease. Morphological research can essentially affect further treatment tactics. If micro metastases without lymph kinesis failure are found patients can remain under dynamic observation. In case of lymph kinesis failure, radical operation on jugular lymph drain ducts is necessary.
O.150 CK 17 in oral squamous cell carcinoma cells by brush biopsy T. Toyoshima, F. Koch, P. Kaemmerer, W. Wagner, B. Al-Nawas. Oral and Maxillofacial Surgery, Johannes Gutenberg University, Mainz, Germany Objectives: Brush biopsy has proved as a non-invasive methodology in diagnosis of oral lesions. Cytokeratin (CK) has been known to be a diagnostic and prognostic marker for solid tumors. The aim of this study was to evaluate the relevance of CKs for diagnosis of oral squamous cell carcinoma (OSCC) using brush biopsy. Methods: Fifty-two pairs of OSCC cells and normal oral mucosal cells were obtained by brush biopsy from OSCC patients. mRNA was extracted from cell pellets for real-time quantitative reverse transcriptase polymerase chain reaction (RT qPCR). The overexpression levels of CK 17, CK 19, and CK 20 in OSCC cells were examined by SYBR green real-time RT qPCR. Results: Compared to in normal mucosal cells, the overexpression of CK 17 was detectable in 40 OSCC cells (76.9%), CK 19 in 19 (36.5%), while that of CK 20 was not detectable in any cells. Compared to the mean value of CK 19 expression level, in all 52 patients, the value of CK 17 was significantly higher (P < 0.02). Moreover, the value of CK 17 was also significantly higher in T1 and T2 OSCC patients (P < 0.03), in patients without metastases of neck lymph nodes (P < 0.02), in stage I and II patients (P < 0.03 and P < 0.05, respectively), and in well differentiated OSCC patients (P < 0.05). Conclusion: The brush biopsy serves for the detection of CK mRNA using real-time RT qPCR. CK 17 could be a promising marker for OSCC diagnosis in brush biopsy test.
O.151 DNA-image cytometry improves assessment of resection margins J. Handschel, C. Sproll, R. Depprich, A. Zimmermann, S. Braunstein, N.R. K¨ubler, U. Meyer, A. B¨ocking. Department for Cranio- and Maxillofacial Surgery, D¨usseldorf, Germany Background: Despite the histopathological findings of tumor free margins, patients with oral squamous cell carcinoma (SCC) often suffer from local tumor relapse. The purpose of this study was to determine the prognostic value of DNA-image cytometry in the assessment of resection margins. Methods: DNA-image cytometry was performed in 40 SCCpatients with histologically tumor-free resection-margins. The follow-up period since the tumor resection was at least 3 years. Results: 20 patients showed a locoregional relapse of the SCC. 14 of these patients had aneuploid cells in DNA-image cytometry. Two patients who were relapse free revealed aneuploid cells too. The sensitivity of the adjuvant use of DNA-image cytometry was 70% and the positive predictive value was 87.5%.
Oral Presentations Conclusions: The additional use of DNA-image cytometry is a reasonable tool for the assessment of the resection margins of SCCs. DNA-image cytometry could help to find the appropriate treatment option for the patients and thus might improve their prognosis. O.152 Endoscopic screening in primary staging of oral cancer M. Kesting, C. Pautke, K.-D. Wolff, T. Muecke. Technical University Munich, Munich, Germany Objectives: Smoking and alcohol abuse are risk factors for oral squamous cell carcinoma (OSCC) as for additional diseases, particularly of the upper gastrointestinal and respiratory tract (UADT). The objective of this study was to assess the relevant data of a single maxillofacial unit. Methods: The clinical records of 806 patients with OSCC between 1997 and 2007 were reviewed. All patients simultaneously underwent esophagogastroduodenoscopy and bronchoscopy. After initial tumor staging patients were classified according to the UICC classification for head and neck malignancies. Clinical and histological data were analyzed for probability by univariate and multivariate analysis. Results: A total of 570 patients (188 female, 382 male) with previously untreated OSCC were identified. 289 patients suffered from limited (stage I and II) and 281 patients from advanced (stage III and IV) disease. A statistically significant positive association between advanced and limited tumor stage (p = 0.038) was found with an increased risk of an additional malignancy for limited tumor stage. Pathologic findings were observed due to esophagogastroduodenoscopy in 169 cases and bronchoscopy in 166 cases. Additional risk factors for the development of secondary UADT cancers were detected in 103 cases. Conclusions: Synchronous secondary diseases of the UADT have a profound impact on survival of patients with OSCC. Esophagogastroduodenoscopy and bronchoscopy have been proven to be effective tools for diagnosis of such diseases and should be included into routinely staging of OSCC. O.153 Genes Ldh-1, Ldh-2, and Ldr-1 in angiogenesis/tumorigenesis
Cancer biology and epidemiology I
S39
by Ldr-1, whose polypeptide, LDR-A, blocks H:M and H:H associations for subsequent angiogenic aerobic energy boosts. With less energy for growth, mice/rat tumors eradicate more easily. Conclusion: LDH data, reported as polypeptides and ratios, offer new evolutionary insights of distinguishable shifts in aerobic status for understanding lagging human clinical trials results. O.154 Host defense peptides in oral cancer M. Kesting, C. Pautke, D. Loeffelbein, F. Hoelzle, T. Muecke, K.-D. Wolff, R. Hasler, La. Steinstraesser. Technical University Munich, Munich, Germany Objectives: Although host defense peptides (HDP) seem to have diverse functional activities in innate antimicrobial immunity, a few reports also indicated an altered expression of these antimicrobial peptides (AMPs) in tissues of cancer. The present study was aimed to screen HDP gene expression in oral squamous cell carcinoma (OSCC) tissue. Methods: During surgical resection tumour tissue and healthy squamous tissue from the contralateral site was harvested from 39 donors using a 4 mm punch-biopsy. After storage, mRNA extraction and cDNA synthesis, tissue was analysed for quantitative LL-37, HBD-1−4, Dermcidin, RNase7 and Psoriasin (S100A7) mRNA expression which was normalized to 18SrRNA as house keeping gene. Additionally fluorescent immunohistochemical staining was performed for HBD-2 and -3, RNase7 and Psoriasin for corresponding localisation of protein expression. Results: The expression of HDPs like Psoriasin, HBD-2+3 or RNase7 was drastically increased within the tumour tissue compared to the healthy one. No differences were observed for LL-37, HBD1 and 4 or Dermcidin. Immunohistochemistry demonstrated specific staining for the investigated peptides in correlation to the expression level of the qRT-PCR experiments. Conclusions: The expression patterns of several HDPs are shown to be significantly altered in OSCCs as compared to intraindividual controls. The present findings may indicate that beside the antimicrobial activity of HDPs, they may also play a role in the pathogenesis of OSCC. O.155 mRNA detection of tumor genes by oral brush biopsy
A. Kunz1 , B. Ahmed2 , C. Stolte3 . 1 Harvard University (Extension), Boston, MA, USA; 2 Department of Surgery, Children’s Hospital, Boston, MA, USA; 3 Broad Institute, MIT, Cambridge, MA, USA
F. Koch, T. Toyoshima, S. Biesterfeld, W. Wagner. Department of Maxillofacial Surgery, University Hospital of Mainz, Mainz, Germany
Introduction: There is a paucity of successful translational data for tumorogenesis in angiogenesis research; while tumors loose their dominant control in mice/rat models, similar results lag in human clinical trials. Methods: This paper explores the evolutionary aspect of 3 genes, Ldh-1, Ldh-2, and Ldr-1. Ldh-1 and Ldh-2 play central roles in regulation of energy supply of all cells; Ldr-1, a regulatory gene, restricted to red blood cells (RBCs) in mice/rats, blocks mammalian random association of LDH-M and LDH-H. Results/Discussion: Remarkable, in evolution, are tissue specific LDH polypeptide ratios, H/M, noting aerobic status, and species specific H/M ratios, meaning, as the evolutionary scale ascends toward man, H/M ratios increase in homologous tissues; striking differences occur in brain and especially angiogenic RBCs. This is important: polypeptides (LDH-H)4 and (LDH-M)4 are LDH-1 and LDH-5 isozymes, respectively, the former yielding 12 times more energy. Species data for rat, monkey, man, respectively: RBC LDH-H: 0%, 64.4% 79%.; RBC H/M: 0.0, 1.8, 3.8; brain LDH-H: 64%, 67%, 70.4%; brain H/M: 1.6, 2.0, 2.7. There are evolutionary increases of LDH-H in RBCs and brain, and evolutionary increases in species aerobic, H/M, status. The lowest H/M value (rats), standard model for angiogenic tumor studies, is controlled
Objectives: The use of reverse transcriptase polymerase chain reaction (RT-PCR) in determining the mRNA of Glutathion-STransferase (GST), Matrixmetalloproteinase (MMP) 2, TenascinC and human Telomerase Reverse Transcriptase (hTERT) was investigated to assist with the diagnosis of Oral Squamous Cell Carcinoma (OSCC) in brush biopsies. Methods: Twenty brush smears of OSCC and the analogous normal mucosa in the same individua were obtained. The cytologic specimens were reviewed. A quantitative RT-PCR for GST, MMP 2, Tenascin C and hTERT mRNA was performed. The mRNA of SCC-cell culture was used as an internal control and relative standard. Results: Compared to normal mucosa GST was higher expressed in 68%, MMP2 in 44%, Tenascin C in 72%, hTERT in 61% of the investigated OSCC cases. The conventional cytology by Haematoxilin & Eosin (HE) stained smears showed a sensitivity of 79%. Three cases had not been diagnosed correctly by conventional brush biopsy, but showed higher mRNA expression of all tested genes compared to normal mucosa. Conclusions: OSCC may be underdiagnosed in conventional brush cytopathology, especially in small, early lesions. The expression of GST, MMP, Tenascin C and hTERT may be a useful diagnostic adjunct in the interpretation of inconclusive cases of
Journal of Cranio-Maxillofacial Surgery 36(2008) Suppl. 1
Tuesday, 9 September 2008, 16.00–17.00
Sala Italia
Cancer biology and epidemiology I O.147 cAMP-phosphodiesterase signal in malignant melanoma cells Y. Watanabe, T. Murata, K. Shimizu, K. Mizoi, J. Nomura, T. Tagawa. Department of Oral and Maxillofacial Surgery, Division of Reparative and Regenerative Medicine, Institute of Medical Science, Mie University Graduate School of Medicine 1, Tsu, Japan Objectives: Cyclic AMP (cAMP) acts as a second messenger in cells and modulates many physiological processes. Intracellular cAMP content is regulated through synthesis by adenylate cyclase and degradation by phosphodiesterases (PDEs). However, no data are available as to the cAMP-PDE signal in malignant melanoma cells. In this study, we examined cAMP-PDE signal in malignant melanoma cell line. Methods: Cell growth: Mouse B16 melanoma cells were plated at 100 cells/well in a 96-well plate and allowed to adhere for 24 h. The cells were cultured in the absence or presence of different concentration of cAMP analog, 8-bromo cAMP, or cGMP analog, 8-bromo cGMP, for 6 days. The cell numbers were calculated using CellTiter 96® Aqueous One Solution Cell Proliferation Assay (Promega, Madison, WI). ˚ cells/25oz. flask. PDE activity: B16 cells were seeded at 1×10A After 3 days, the cells were incubated with 8-bromo cAMP for 15 min, harvested and homogenized in 1 ml of ice-cold homogenization buffer. PDE activity was measured. Results: 8-bromo cAMP inhibited the growth of B16 cells, but not 8-bromo cGMP. And PDE activity in B16 cells was stimulated by 8-bromo cAMP. Conclusions: These data suggested that cAMP-PDE singal was related with the growth of malignant melanoma cells and cAMPPDE singal might be target for an antitumor drug of malignant melanoma. O.148 Chemical surgery for oral cancer S. Kim. Sacred Heart Hospital, Anyang, Republic of Korea Objective: The control of microinvasion is highly important to improve complete cure rate in oral cancer. We wanted to evaluate the therapeutic effects of chemical agent that has sharp threshold in inducing apoptosis of cancer. Methods: Eight patients were included in this clinical trials and six patients might have remained cancer in resection margin. Topical 4-HR application was done for 1 month postoperatively. Results: Seven patients showed tumor free and mean follow-up was 32 months. There was no metastatic tumor after operation. Conclusion: Though this was a preliminary trials, the chemical surgery could be considered as an additive technique to improve cure rate of oral cancer. O.149 Choice of operation volume in glossal cancer T1-3NxMO R. Bakirov, S. Osokin, R. Osokina. Bashkir Clinical Oncology Health Center, UFA, Russia The aim of our research is to evaluate effectiveness of lymph node dissection as an approach which allows giving prognosis of metastasizing in remote postoperative period. We analyzed history cases of 256 patients with glossal cancer T1−3NxMo who received surgical treatment in Head and Neck Tumor Department. We found out that patients who received treatment only within gemiglossectomy had regional metastases in jugular lymph nodes in 28% of cases.
Abstracts, EACMFS XIX Congress The second group of patients is those who received treatment of initial lesion having simultaneous removing of regional lymph drain ducts whose metastasizing process wasn’t proved by the results of diagnostic study. In this group of patients the frequency of metastases to remote lymph nodes was 23%. In 88.4% of operated patients of this group there were no metastases within one year. The patients of the third group in 10.5% of cases had metastases within one year after operation. Simultaneous regional lymph node dissection and operation of the initial lesion T1−3NxMo can be treated as a factor of surgical prognosis of further course of disease. Morphological research can essentially affect further treatment tactics. If micro metastases without lymph kinesis failure are found patients can remain under dynamic observation. In case of lymph kinesis failure, radical operation on jugular lymph drain ducts is necessary.
O.150 CK 17 in oral squamous cell carcinoma cells by brush biopsy T. Toyoshima, F. Koch, P. Kaemmerer, W. Wagner, B. Al-Nawas. Oral and Maxillofacial Surgery, Johannes Gutenberg University, Mainz, Germany Objectives: Brush biopsy has proved as a non-invasive methodology in diagnosis of oral lesions. Cytokeratin (CK) has been known to be a diagnostic and prognostic marker for solid tumors. The aim of this study was to evaluate the relevance of CKs for diagnosis of oral squamous cell carcinoma (OSCC) using brush biopsy. Methods: Fifty-two pairs of OSCC cells and normal oral mucosal cells were obtained by brush biopsy from OSCC patients. mRNA was extracted from cell pellets for real-time quantitative reverse transcriptase polymerase chain reaction (RT qPCR). The overexpression levels of CK 17, CK 19, and CK 20 in OSCC cells were examined by SYBR green real-time RT qPCR. Results: Compared to in normal mucosal cells, the overexpression of CK 17 was detectable in 40 OSCC cells (76.9%), CK 19 in 19 (36.5%), while that of CK 20 was not detectable in any cells. Compared to the mean value of CK 19 expression level, in all 52 patients, the value of CK 17 was significantly higher (P < 0.02). Moreover, the value of CK 17 was also significantly higher in T1 and T2 OSCC patients (P < 0.03), in patients without metastases of neck lymph nodes (P < 0.02), in stage I and II patients (P < 0.03 and P < 0.05, respectively), and in well differentiated OSCC patients (P < 0.05). Conclusion: The brush biopsy serves for the detection of CK mRNA using real-time RT qPCR. CK 17 could be a promising marker for OSCC diagnosis in brush biopsy test.
O.151 DNA-image cytometry improves assessment of resection margins J. Handschel, C. Sproll, R. Depprich, A. Zimmermann, S. Braunstein, N.R. K¨ubler, U. Meyer, A. B¨ocking. Department for Cranio- and Maxillofacial Surgery, D¨usseldorf, Germany Background: Despite the histopathological findings of tumor free margins, patients with oral squamous cell carcinoma (SCC) often suffer from local tumor relapse. The purpose of this study was to determine the prognostic value of DNA-image cytometry in the assessment of resection margins. Methods: DNA-image cytometry was performed in 40 SCCpatients with histologically tumor-free resection-margins. The follow-up period since the tumor resection was at least 3 years. Results: 20 patients showed a locoregional relapse of the SCC. 14 of these patients had aneuploid cells in DNA-image cytometry. Two patients who were relapse free revealed aneuploid cells too. The sensitivity of the adjuvant use of DNA-image cytometry was 70% and the positive predictive value was 87.5%.
Oral Presentations Conclusions: The additional use of DNA-image cytometry is a reasonable tool for the assessment of the resection margins of SCCs. DNA-image cytometry could help to find the appropriate treatment option for the patients and thus might improve their prognosis. O.152 Endoscopic screening in primary staging of oral cancer M. Kesting, C. Pautke, K.-D. Wolff, T. Muecke. Technical University Munich, Munich, Germany Objectives: Smoking and alcohol abuse are risk factors for oral squamous cell carcinoma (OSCC) as for additional diseases, particularly of the upper gastrointestinal and respiratory tract (UADT). The objective of this study was to assess the relevant data of a single maxillofacial unit. Methods: The clinical records of 806 patients with OSCC between 1997 and 2007 were reviewed. All patients simultaneously underwent esophagogastroduodenoscopy and bronchoscopy. After initial tumor staging patients were classified according to the UICC classification for head and neck malignancies. Clinical and histological data were analyzed for probability by univariate and multivariate analysis. Results: A total of 570 patients (188 female, 382 male) with previously untreated OSCC were identified. 289 patients suffered from limited (stage I and II) and 281 patients from advanced (stage III and IV) disease. A statistically significant positive association between advanced and limited tumor stage (p = 0.038) was found with an increased risk of an additional malignancy for limited tumor stage. Pathologic findings were observed due to esophagogastroduodenoscopy in 169 cases and bronchoscopy in 166 cases. Additional risk factors for the development of secondary UADT cancers were detected in 103 cases. Conclusions: Synchronous secondary diseases of the UADT have a profound impact on survival of patients with OSCC. Esophagogastroduodenoscopy and bronchoscopy have been proven to be effective tools for diagnosis of such diseases and should be included into routinely staging of OSCC. O.153 Genes Ldh-1, Ldh-2, and Ldr-1 in angiogenesis/tumorigenesis
Cancer biology and epidemiology I
S39
by Ldr-1, whose polypeptide, LDR-A, blocks H:M and H:H associations for subsequent angiogenic aerobic energy boosts. With less energy for growth, mice/rat tumors eradicate more easily. Conclusion: LDH data, reported as polypeptides and ratios, offer new evolutionary insights of distinguishable shifts in aerobic status for understanding lagging human clinical trials results. O.154 Host defense peptides in oral cancer M. Kesting, C. Pautke, D. Loeffelbein, F. Hoelzle, T. Muecke, K.-D. Wolff, R. Hasler, La. Steinstraesser. Technical University Munich, Munich, Germany Objectives: Although host defense peptides (HDP) seem to have diverse functional activities in innate antimicrobial immunity, a few reports also indicated an altered expression of these antimicrobial peptides (AMPs) in tissues of cancer. The present study was aimed to screen HDP gene expression in oral squamous cell carcinoma (OSCC) tissue. Methods: During surgical resection tumour tissue and healthy squamous tissue from the contralateral site was harvested from 39 donors using a 4 mm punch-biopsy. After storage, mRNA extraction and cDNA synthesis, tissue was analysed for quantitative LL-37, HBD-1−4, Dermcidin, RNase7 and Psoriasin (S100A7) mRNA expression which was normalized to 18SrRNA as house keeping gene. Additionally fluorescent immunohistochemical staining was performed for HBD-2 and -3, RNase7 and Psoriasin for corresponding localisation of protein expression. Results: The expression of HDPs like Psoriasin, HBD-2+3 or RNase7 was drastically increased within the tumour tissue compared to the healthy one. No differences were observed for LL-37, HBD1 and 4 or Dermcidin. Immunohistochemistry demonstrated specific staining for the investigated peptides in correlation to the expression level of the qRT-PCR experiments. Conclusions: The expression patterns of several HDPs are shown to be significantly altered in OSCCs as compared to intraindividual controls. The present findings may indicate that beside the antimicrobial activity of HDPs, they may also play a role in the pathogenesis of OSCC. O.155 mRNA detection of tumor genes by oral brush biopsy
A. Kunz1 , B. Ahmed2 , C. Stolte3 . 1 Harvard University (Extension), Boston, MA, USA; 2 Department of Surgery, Children’s Hospital, Boston, MA, USA; 3 Broad Institute, MIT, Cambridge, MA, USA
F. Koch, T. Toyoshima, S. Biesterfeld, W. Wagner. Department of Maxillofacial Surgery, University Hospital of Mainz, Mainz, Germany
Introduction: There is a paucity of successful translational data for tumorogenesis in angiogenesis research; while tumors loose their dominant control in mice/rat models, similar results lag in human clinical trials. Methods: This paper explores the evolutionary aspect of 3 genes, Ldh-1, Ldh-2, and Ldr-1. Ldh-1 and Ldh-2 play central roles in regulation of energy supply of all cells; Ldr-1, a regulatory gene, restricted to red blood cells (RBCs) in mice/rats, blocks mammalian random association of LDH-M and LDH-H. Results/Discussion: Remarkable, in evolution, are tissue specific LDH polypeptide ratios, H/M, noting aerobic status, and species specific H/M ratios, meaning, as the evolutionary scale ascends toward man, H/M ratios increase in homologous tissues; striking differences occur in brain and especially angiogenic RBCs. This is important: polypeptides (LDH-H)4 and (LDH-M)4 are LDH-1 and LDH-5 isozymes, respectively, the former yielding 12 times more energy. Species data for rat, monkey, man, respectively: RBC LDH-H: 0%, 64.4% 79%.; RBC H/M: 0.0, 1.8, 3.8; brain LDH-H: 64%, 67%, 70.4%; brain H/M: 1.6, 2.0, 2.7. There are evolutionary increases of LDH-H in RBCs and brain, and evolutionary increases in species aerobic, H/M, status. The lowest H/M value (rats), standard model for angiogenic tumor studies, is controlled
Objectives: The use of reverse transcriptase polymerase chain reaction (RT-PCR) in determining the mRNA of Glutathion-STransferase (GST), Matrixmetalloproteinase (MMP) 2, TenascinC and human Telomerase Reverse Transcriptase (hTERT) was investigated to assist with the diagnosis of Oral Squamous Cell Carcinoma (OSCC) in brush biopsies. Methods: Twenty brush smears of OSCC and the analogous normal mucosa in the same individua were obtained. The cytologic specimens were reviewed. A quantitative RT-PCR for GST, MMP 2, Tenascin C and hTERT mRNA was performed. The mRNA of SCC-cell culture was used as an internal control and relative standard. Results: Compared to normal mucosa GST was higher expressed in 68%, MMP2 in 44%, Tenascin C in 72%, hTERT in 61% of the investigated OSCC cases. The conventional cytology by Haematoxilin & Eosin (HE) stained smears showed a sensitivity of 79%. Three cases had not been diagnosed correctly by conventional brush biopsy, but showed higher mRNA expression of all tested genes compared to normal mucosa. Conclusions: OSCC may be underdiagnosed in conventional brush cytopathology, especially in small, early lesions. The expression of GST, MMP, Tenascin C and hTERT may be a useful diagnostic adjunct in the interpretation of inconclusive cases of