Marcadores moleculares en pacientes con cáncer de cuello uterino tratadas con quimioterapia neoadyuvante y radioterapia / Molecular markers in patients with cervical cancer treated with induction chemotherapy and radiation therapy

Objetivos: En este estudio prospectivo de determinaron las modificaciones en la expresión y el valor predictivo de p53, p21 wafi/sdII/cipi, PCNA, hMLH1, hMSH2, Bcl'2 y TUNEL en pacientes con cáncer de cervix localmente avanzado tratadas con quimioterapia de inducción y radioterapia. Pacientes y métodos: Se obtuvieron muestras de 24 pacientes (IB'bulky/IIIB, 95 por ciento carcinomas escamosos) antes de la quimioterapia y a los 30 días del tratamiento. Trece pacientes recibieron un esquema de drogas basado en cisplatino y como la respuesta a esta terapia no fue buena, a las otras 11 pacientes se les administró vinorelbine e ifosfamida. Luego de la quimioterapia todas las pacientes recibieron radioterapia. La expresión de los marcadores moleculares en las biopsias pre- y post quimioterapia se estudió por inmunohistoquímica y la apoptosis fue evaluada por la técnica del TUNEL mejorada recientemente. Para comparar los cambios en la expresión de los marcadores moleculares y para correlacionarlos con la evaluación clínica (media de seguimiento: 31 meses para las pacientes que recibieron cisplatino y 19 para las que recibieron vinorelbine e ifosfamida) se realizaron análisis estadísticos. Resultados y conclusiones: La quimioterapia de inducción no aumentó la sobrevida de las pacientes, el 50 por ciento tuvo enfermedad progresiva (EP) o falleció (F). La expresión de p21waf1/sdII/cip1, hMLF1, hMSH2, y Bcl-2 no mostró cambios significativos después de la quimioterapia y no correlacionó con la evaluación clínica. La expresión de p53 no se modificó luego de la quimioterapia, las pacientes con tumores p53 positivos mostraron una tendencia a tener una sobrevida menor. Las pacientes con EP o que fallecieron mostraron niveles altos de PCNA, a diferencia de aquellas que estuvieron libres de enfermedad (LE) o con enfermedad estable (EE) (50 por ciento versus 17 por ciento, respectivamente, p<0.004). La sobrevida de las pacientes con bajos índices de TUNEL (igual o menor al valor medio entre las biopsias pre y post-quimioterapia de 1.5) fue significativamente más corta que las pacientes que presentaron índices de TUNEL altos (p<0.009). Nuestros resultados muestran que la quimioterapia de inducción (los dos tratamientos aplicados en este estudio) no mejoró la sobrevida de pacientes con cáncer de cervix...

Marcadores moleculares en pacientes con cáncer de cuello uterino tratadas con quimioterapia neoadyuvante y radioterapia / Molecular markers in patients with cervical cancer treated with induction chemotherapy and radiation therapy

Objetivos: En este estudio prospectivo de determinaron las modificaciones en la expresión y el valor predictivo de p53, p21 wafi/sdII/cipi, PCNA, hMLH1, hMSH2, Bcl2 y TUNEL en pacientes con cáncer de cervix localmente avanzado tratadas con quimioterapia de inducción y radioterapia. Pacientes y métodos: Se obtuvieron muestras de 24 pacientes (IBbulky/IIIB, 95 por ciento carcinomas escamosos) antes de la quimioterapia y a los 30 días del tratamiento. Trece pacientes recibieron un esquema de drogas basado en cisplatino y como la respuesta a esta terapia no fue buena, a las otras 11 pacientes se les administró vinorelbine e ifosfamida. Luego de la quimioterapia todas las pacientes recibieron radioterapia. La expresión de los marcadores moleculares en las biopsias pre- y post quimioterapia se estudió por inmunohistoquímica y la apoptosis fue evaluada por la técnica del TUNEL mejorada recientemente. Para comparar los cambios en la expresión de los marcadores moleculares y para correlacionarlos con la evaluación clínica (media de seguimiento: 31 meses para las pacientes que recibieron cisplatino y 19 para las que recibieron vinorelbine e ifosfamida) se realizaron análisis estadísticos. Resultados y conclusiones: La quimioterapia de inducción no aumentó la sobrevida de las pacientes, el 50 por ciento tuvo enfermedad progresiva (EP) o falleció (F). La expresión de p21waf1/sdII/cip1, hMLF1, hMSH2, y Bcl-2 no mostró cambios significativos después de la quimioterapia y no correlacionó con la evaluación clínica. La expresión de p53 no se modificó luego de la quimioterapia, las pacientes con tumores p53 positivos mostraron una tendencia a tener una sobrevida menor. Las pacientes con EP o que fallecieron mostraron niveles altos de PCNA, a diferencia de aquellas que estuvieron libres de enfermedad (LE) o con enfermedad estable (EE) (50 por ciento versus 17 por ciento, respectivamente, p<0.004). La sobrevida de las pacientes con bajos índices de TUNEL (igual o menor al valor medio entre las biopsias pre y post-quimioterapia de 1.5) fue significativamente más corta que las pacientes que presentaron índices de TUNEL altos (p<0.009). Nuestros resultados muestran que la quimioterapia de inducción (los dos tratamientos aplicados en este estudio) no mejoró la sobrevida de pacientes con cáncer de cervix... (AU)

Effects of short-term tamoxifen administration in patients with invasive cervical carcinoma.

Cervical cancer is not considered a hormone-responsive tumor in spite of the presence of estrogen receptors (ER) and progesterone receptors (PgR) in some of them. Endocrine treatments have not achieved clinical responses, however, tamoxifen has been reported to induce PgR and to inhibit cell growth of many cervical carcinoma cell lines. In this study we investigated whether tamoxifen administration affects the histopathological characteristics of cervical cancer and the expression of ER, PgR, HER-2/neu and p53 protein. Nineteen patients with invasive cervical cancer free of previous treatments were studied. The triphenylethylene antiestrogen tamoxifen was given orally during 10 days (20 or 40 mg/day). Pre- and post-tamoxifen biopsies were evaluated using slides stained with hematoxylin and eosin and immunostained (ER, PgR, HER-2/neu, p53, PCNA, keratin, heat shock protein 27,000 daltons). Estrogen receptors were present in 37% and PgR in 16% of the biopsies from untreated patients. Only one case that was PgR-negative before tamoxifen administration showed weak PgR-positivity following antiestrogen administration. No obvious changes were observed in ER, HER-2/neu and p53 proteins. A statistically significant decrease in the number of mitotic figures was obtained in 16% (3/19) of the post-tamoxifen biopsies and two of them showed higher differentiation. The results showed that tamoxifen did not induce changes in estrogen-regulated proteins in cervical cancer. However, the data showed that certain cervical carcinomas had changes in their proliferation and differentiation levels following tamoxifen administration. These findings suggest that tamoxifen may affect some cervical cancer tissues by a hormone-independent mechanism(s).

Effect of human papillomavirus infection on estrogen receptor and heat shock protein hsp27 phenotype in human cervix and vagina.

In this study we have explored whether, as a consequence of human papillomavirus (HPV) infection, there is inappropriate expression of estrogen receptor and/or of a small heat shock protein of 27,000 daltons (hsp27). Estrogen receptor, hsp27, and HPV structural antigens were detected by immunocytochemistry, while HPV DNA (6/11, 16/18, 31/35/51) was determined by in situ hybridization in cervical and vaginal samples from 40 patients. Most of the samples with HPV infection without atypia showed a shift in estrogen receptor expression since this protein appeared mainly in the intermediate and superficial cell layers. In the serial sections, these layers displayed strong estrogen receptor staining, together with high HPV replication and late HPV gene expression. In the samples with HPV infection and atypia, estrogen receptors were also frequently found in the basal and parabasal cells, but almost 20% of these samples did not show estrogen receptors. The presence of high estrogen receptor expression was not dependent on a particular HPV DNA type. On the other hand, interesting modifications in hsp27 expression were observed in the HPV-infected tissues. The cytoplasm of the cells with koilocytotic changes showed very low hsp27 content. In several samples this protein appeared in the nuclei of the superficial cells, and sometimes it was also observed in the cytoplasm of the basal cells. These changes in estrogen receptor and hsp27 expression suggest that these proteins might have a role in virus-host biology.

Vagal stimulation and exogenous acetylcholine in isolated rat stomach: calcitonin effect on contractile activity.

Calcitonin induces contraction in the vascular and extravascular smooth muscle and facilitates the transmission of the excitation in somatic motor nerve endings. These actions are Ca2(+)-dependent. The calcitonin effect on autonomic nerve endings has been studied here by testing influence of calcitonin on the contractile responses of the isolated rat stomach. The organ was submitted to electrical vagal stimulation or, after denervation, to exogenous acetylcholine. Calcitonin invariably increased the muscular tone and reduced the contractile responses to vagal stimulation. Opposite effects were noted after a serotoninergic block with nicergoline. Calcitonin also increased the contractile response evoked by exogenous acetylcholine and the Ca(2)-antagonist nicardipine counteracted the facilitatory effects. We suggest that the inhibitory action of calcitonin is serotonine-dependent while the facilitatory one is Ca2(+)-dependent.

Diode laser in cervical myofascial pain: a double-blind study versus placebo.

We present a double-blind trial in which a pulsed infrared beam was compared with a placebo in the treatment of myofascial pain in the cervical region. The patients were submitted to 12 sessions on alternate days to a total energy dose of 5 J each. At each session, the four most painful muscular trigger points and five bilateral homometameric acupuncture points were irradiated. Those in the placebo group submitted to the same number of sessions following an identical procedure, the only difference being that the laser apparatus was nonoperational. Pain was monitored using the Italian version of the McGill pain questionnaire and the Scott-Huskisson visual analogue scale. The results show a pain attenuation in the treated group and a statistically significant difference between the two groups of patients, both at the end of therapy and at the 3-month follow-up examination.

Analysis of a 24-kilodalton (KD) protein in the human uterine cervix during abnormal growth.

The authors have previously studied the presence and distribution of a 24-kilodalton (KD) estrogen-regulated protein in the human normal cervix (Am J Obstet Gynecol 1986; 155:1090-1096). This protein has recently been identified as a heat-shock protein, and in order to continue its study the authors have now examined its expression in preneoplastic to neoplastic cervical samples. The study involved 53 patients, the presence of 24-KD protein together with keratin and carcinoembryonic antigen (CEA) was investigated by immunohistochemical analysis. Cytosol samples from 15 patients with squamous cervical carcinomas were also studied by the Western blot technique, and the presence of estrogen receptors was analyzed biochemically. The 24-KD protein was observed in cervical intraepithelial neoplasias (CIN), but it was not useful to identify the different degrees of CIN examined. The 24-KD protein, keratin, and CEA were predominantly expressed in well and moderately differentiated squamous carcinomas in the more differentiated areas, and the protein was also found in cervical adenocarcinomas. The presence of 24-KD protein did not correlate with that of estrogen receptors in squamous cervical carcinomas. The Western blot and the immunohistochemical studies revealed that the antibody to 24-KD protein does not cross-react with epitopes of CEA and keratins.

Pharmacokinetic studies with zinc(II)-phthalocyanine in tumour-bearing mice.

Zn(II)-phthalocyanine (Zn-Pc) incorporated into unilamellar liposomes of dipalmitoylphosphatidylcholine has been injected intraperitoneally (0.5 mg kg-1) to BALB/c mice bearing a transplanted MS-2 fibrosarcoma. The drug is specifically transported by serum lipoproteins and cleared from the serum via the bile-gut pathway in a biphasic process: approximately 60% of Zn-Pc is eliminated with a serum half-life of approximately 9 hours, while the remaining aliquot is eliminated at a very slow rate. Several normal tissues take up the drug within 3 hours after administration but release it almost completely after 24-48 hours. On the other hand, the tumour shows a maximum concentration of Zn-Pc (approximately 0.6 microgram g-1 of tissue) after 18-24 hours; at this time, the ratio between the Zn-Pc levels in the tumour and the muscle (which represents the surrounding normal tissue) is approximately 7.5. The results are discussed in terms of a possible use of Zn-Pc as a photosensitizer in the photodynamic therapy of tumours.

Coexistencia de lesiones blancas de cervix y vulva correlación histológica y rol del virus papiloma / Coexistance of white cervix and vulga injuries histological correlation and papillomavirus

Se estudió la incidencia de la infección por Papiloma virus en 300 pacientes ambulatorias, de las cuales 120 presentaron algún signo de infección viral. El 52% eran lesiones multicéntricas y simultáneas en cérvix y vulva. El 10% de las pacientes con colposcopia negativa tenían signos citológicos de infección viral. La vulva, resultó ser un "órgano target" muy frecuente en este agente infeccioso (77%) y que en muchos casos precede a la lesión cervical. Se recalca la importancia del examen de toda la región anogenital como sustrato posible del desarrollo de una lesión viral de alto potencial oncogénico

Coexistencia de lesiones blancas de cervix y vulva correlación histológica y rol del virus papiloma / Coexistance of white cervix and vulga injuries histological correlation and papillomavirus

Se estudió la incidencia de la infección por Papiloma virus en 300 pacientes ambulatorias, de las cuales 120 presentaron algún signo de infección viral. El 52% eran lesiones multicéntricas y simultáneas en cérvix y vulva. El 10% de las pacientes con colposcopia negativa tenían signos citológicos de infección viral. La vulva, resultó ser un "órgano target" muy frecuente en este agente infeccioso (77%) y que en muchos casos precede a la lesión cervical. Se recalca la importancia del examen de toda la región anogenital como sustrato posible del desarrollo de una lesión viral de alto potencial oncogénico (AU)

Localization of an estrogen-responsive protein in the human cervix during menstrual cycle, pregnancy, and menopause and in abnormal cervical epithelia without atypia.

The presence and distribution of an estrogen-responsive protein with a molecular weight of 28,000 were investigated in the human cervix with use of a monoclonal antibody. This biochemical marker protein was localized with a light microscope and by immunocytochemical studies of the different cell types and cell layers of the cervix. The study involved 60 patients, 48 of whom were sexually active, eight pregnant, and four in menopause; strips of endometrial tissue were analyzed in 22 patients. In cycling women the estrogen-responsive protein was identified in the subcolumnar cells of the endocervix, whereas in the ectocervix the protein was detected mainly in the parabasal and intermediate cell layers but alternating with unstained areas. There were no significant variations in the presence of the protein in the ectocervical and endocervical epithelium during the different phases of the menstrual cycle. During pregnancy a more intense and homogeneous immunostaining of the protein was seen in the ectocervix, endocervical areas with squamous metaplasia showed strong estrogen-responsive immunostaining, and predecidual and decidual cells were positive for the protein. There were no prominent changes in the presence and distribution of the protein in the abnormal ectocervical samples without atypia. However, in the endocervix the protein detection was useful to follow the evolution of the subcolumnar cells to simple squamous metaplasia. These samples displayed intense estrogen-responsive immunostaining. No immunoreaction was observed in the cervix of menopausal women. The results of the present study have shown that the response of the normal uterine cervix to estrogenic influence is heterogeneous in different cervical cell types and in different sites within the same cell layer; during the normal menstrual cycle the capability of response of the cervical cells to variations of estrogen levels is limited when compared with the endometrium; and during pregnancy and in the process of indirect squamous metaplasia some of the cervical cells seem to be very reactive to estrogenic stimulation. This study defines the normal baseline for further analysis of the estrogen-regulated protein in the uterine cervix during abnormal growth.

Sustrato histologico del estudio colposcopico y citologico de 3.078 casos / Histological substrate of colposcopic and cytologic study of the 3.078 casos

Se estudiaron 3.078 pacientes desde el punto de vista colpo-cito-histologia. Fueron biopsadas 205 pacientes por presentar colposcopia sospechosa en imagenes unicas o multiples por lo cual el numero de biopsias se elevo a 254. Las lesiones colposcopicas mas frecuentes fueron mosaico y base. El hallazgo histologico mas frecuente fue la cervicitis cronica y la displasia leve. El indice de NIC III (displasia severa y CIS) fue del 3,25%. Se destacon las virtudes del metodo en la deteccion precoz del cancer cervico uterino

Sustrato histologico del estudio colposcopico y citologico de 3.078 casos / Histological substrate of colposcopic and cytologic study of the 3.078 casos

Se estudiaron 3.078 pacientes desde el punto de vista colpo-cito-histologia. Fueron biopsadas 205 pacientes por presentar colposcopia sospechosa en imagenes unicas o multiples por lo cual el numero de biopsias se elevo a 254. Las lesiones colposcopicas mas frecuentes fueron mosaico y base. El hallazgo histologico mas frecuente fue la cervicitis cronica y la displasia leve. El indice de NIC III (displasia severa y CIS) fue del 3,25%. Se destacon las virtudes del metodo en la deteccion precoz del cancer cervico uterino