ABSTRACT
El virus papiloma humano (VPH) es el principal factor causal del cáncer cervicouterino (CCU). Así, detectar y genotipificar el VPH es importante para conocer la frecuencia de los genotipos presentes en la región. En este trabajo se estudiaron 44 biopsias de adenocarcinoma cervical (ACC). Para la detección del VPH se empleó una reacción de polimerasa en cadena anidada dirigida al gen L1 (RPCL1), para la genotipificación viral se utilizaron enzimas de restricción (Rsa I, Dde I, Pst I) y secuenciación. Se detectó ADN viral mediante RPCL1 anidada en 100 por ciento de las biopias. Se logró tipificar 38/44 casos: 81,6 por ciento VPH 16; 13,2 por ciento VPH 18; 2,6 por ciento VPH 33 y 2,6 por ciento VPH 18/33. Conclusiones: La metodología fue exitosa para identificar el tipo viral en 86 por ciento de las biopsias. Se observó una estrecha asociación ACC-VPH, especialmente con el tipo viral 16, detectado en 81,6 por ciento de los casos tipificados.
Human papillomavirus (HPV) is the main cause of cervical cancer. Thus, HPV detection and typing becomes important in order to know the frequency of genotypes present in the region. In this paper we studied 44 biopsies of cervical adenocarcinoma. For HPV detection nested polymerase chain reaction (PCR) was used to amplify the L1 gene. For viral typing restriction enzymes (Rsa I, Dde I, Pst I) and DNA sequencing were used. Viral DNA was detected by nested L1 PCR in 100 percent of biopsies; 38/44 cases could be typed: 81.6 percent HPV16; 13.2 percent HPV 18; 2.6 percent VPH 33 and 2.6 percent HPV 18/33. Conclusions: The technique was successful in identifying the virus type in 86 percent of biopsies. There was a strong association ACC-HPV, especially with the viral type 16, detected in 81.6 percent of established cases.
Subject(s)
Adult , Aged , Female , Humans , Middle Aged , Adenocarcinoma/virology , Alphapapillomavirus/genetics , DNA, Viral/analysis , Papillomavirus Infections/virology , Uterine Cervical Neoplasms/virology , Chile , Genotype , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Retrospective StudiesABSTRACT
Background: There is a very strong documented correlation between the appearance of cancer cells in blood and occurrence of metastasis in gastrointestinal cancer. Aim: To determine MUC1, CK19, CK20 and CEA mRNA expression in bone marrow of patients with gallbladder cancer and evaluate its clinical significance. Material and methods: Sixty eight samples were analyzed, 38 bone marrow samples of gallbladder cancer patients, 20 healthy donors, and 10 frozen samples of gallbladder cancer. Nested reverse transcriptase-polymerase chain reaction (nested RT-PCR) was used to analyze mRNA expression. Results: All frozen tumors were positive for CEA, CK19, and MUC1 mRNA and 70 percent were positive for CK20. Seventeen of 20 donor samples were positive for MUC1 and only one sample from donors was positive for both CK20 and CK19 mRNA. Among the 38 blood and bone marrow samples of gallbladder cancer patients, the expression of MUC1, CK19, CK20, and CEA, mRNA was 60.5 percent (23/38), 31.6 percent (12/38), 7.9 percent (3/38), and 7.9 percent (3/38), respectively. Disregarding the MUC1 results. 37 percent (14/38), 13 percent (5/38) and 5 percent (2/38) were positive for one, two and three markers respectively. Not significant differences were found in survival with a follow up to 12 months. Conclusion: Our results indicate that the molecular detection of tumor cells in bone marrow in patients with gallbladder carcinoma is technically possible, being CEA, CK19 and CK20 gene expression the best markers. The MUC1 gene expression marker was highly unspecific and it should not been considered. The detection of bone marrow micrometastasis might be helpful in prognosis and the selection of clinical treatment but a larger series with a longer follow-up should be studied.