Extra-pancreatic expression of the rat islet amyloid polypeptide (amylin) gene.

Messenger RNA for rat islet amyloid polypeptide (IAPP) has been identified not only in the pancreas but also, in lesser amounts, in preparations from the stomach and dorsal root ganglia. In the stomach, insulin mRNA was not detectable, ruling out possible contamination by pancreatic tissue. Because IAPP and calcitonin gene-related peptide (CGRP) are related and CGRP is present in both stomach and dorsal root ganglia, it was possible that 'IAPP' signals were in fact due to cross-hybridization with CGRP mRNA. A second IAPP probe was constructed which does not cross-react. This probe also detected mRNA in both tissues, confirming the expression of IAPP in both tissues. The regional distribution of IAPP mRNA in the stomach did not parallel that of gastrin mRNA. IAPP mRNA was present in the antrum, centrum and pylorus and, like gastrin, the highest amounts were in the pylorus. However, the ratio between the pylorus and centrum was 3.6:1 for IAPP and 156:1 for gastrin. The effects of dietary manipulation were determined; a period of 48 h of starvation reduced pancreatic IAPP mRNA by approximately 60%, whereas in the stomach there was no significant reduction. If the action of IAPP was hormonal, pancreas and stomach would not be acting in concert. A paracrine role for gastric IAPP therefore seems more likely.

Chromosomal aberrations in young cancer patients.

Spontaneous level of chromosomal aberrations (CA) is considered to be indicative of inherent cancer predisposition, which plays a major role in total cancer incidence. We have studied spontaneous CA levels in in vitro cultured peripheral blood lymphocytes of pediatric cancer patients (n = 77). Results were compared with those of control subjects (n = 72), including: age-matched controls; elder controls (minimum age 60 years); and healthy first-degree relatives (FDR) of pediatric cancer patients. Pediatric cancer patients showed the highest mean CA/cell value, which was statistically significant as compared to their age-matched counterparts, elder controls, and the FDRs. As compared to 7% of all the three control groups collectively, 32.4% of pediatric cancer patients showed > 0.1 mean CA/cell value. One of the FDRs with a very high frequency of CA developed cancer within three years. The results suggest that spontaneous levels of chromosomal aberrations may be used as one of the biomarkers for cancer predisposition study.

Spontaneous chromosomal instability in breast cancer families.

Spontaneous chromosomal instability has been correlated with cancer predisposition. In the present study, the phenomenon has been evaluated using two cytogenetic markers, namely, frequency of spontaneous sister chromatid exchanges (SCE) and spontaneous chromosomal aberrations (CA) in peripheral blood lymphocytes of hereditary breast cancer (HBC) patients (n = 11) and healthy blood relatives (HBR, n = 36). A statistically significant difference was observed for both the endpoints between HBC patients and controls (P < 0.001), HBC patients and HBR (P < 0.001), as well as HBR and controls (P < 0.001). Thus, 63.64% of the HBC patients and 25% of HBR showed a mean CA/cell value higher than the highest mean CA/cell value of the controls (0.11 CA/cell). Similarly, 81.81% of the HBC patients and 61.11% of HBR showed a mean SCE/cell value higher than the highest mean SCE/cell value of the controls (9.60 SCE/cell). Chromosomal aberrations were more frequently observed in the B and E group of chromosomes in HBC patients and HBR. These findings primarily indicate the high level of chromosomal instability in breast cancer families, and might be one of the predisposing factors for high risk of cancer in HBR.

Circulating prolactin and tumoral prolactin receptors in men with tongue cancer: a preliminary study.

Pre-therapeutic circulating prolactin levels and tumoral prolactin receptors (PRLR) were determined in 25 male patients with tongue cancer. The patients were divided into PRLR positive (PRLR+) and PRLR negative (PRLR-) groups. The overall survival was statistically non-significant between these two groups of PRLR as well as when considering 2% as the cut-off level. Moreover, no correlation was observed between PRLR status and clinicopathologic prognosticators. Furthermore, patients with < 2% PRLR had significantly higher levels of circulating prolactin than their counterparts (P < 0.05). Patients with PRLR- tumors having hyperprolactinemia (prolactin > 15.0 ng/ml) had unfavourable overall survival (chi 2 = 4.08, df = 1, P < 0.04) than those with normoprolactin (prolactin < 15.0 ng/ml). From this pilot study, it seems that PRLR negativity with hyperprolactinemia could be used as an independent predictor of short-term prognosis in cancer tongue patients.

Prolactin as a local growth promoter in patients with breast cancer: GCRI experience.

AIMS: The aim of this study was to evaluate the prognostic value of pre-operative prolactin (PRL) in conjunction with established prognosticators, and the risk of disease relapse in patients with early and advanced breast cancer. To confirm the hypothesis that PRL is produced by breast tumours molecular analysis of PRL, using immunohistochemistry, mRNA by RT-PCR and direct sequencing, was performed. Furthermore, presence of prolactin receptors (PRLR) was evaluated by immunohistochemical localization in these patients. METHODS: In 111 breast cancer patients, pre-operative PRL was determined by an immunoradiometric assay (IRMA) method. Immunohistochemical localization of PRL (IHL-PRL) and PRLR was performed on formalin-fixed, paraffin-embedded tissue sections. Expression of PRL mRNA was carried out by reverse transcriptase polymerase chain reaction (RT-PCR). RT-PCR PRL amplimer was sequenced and compared with human pituitary PRL amplimer. RESULTS: Fifty-eight per cent (64/111) of the patients had hyperprolactinaemia (PRL520.0 ng/ml). With increasing tumour size, a higher incidence of hyperprolactinaemia was noted which was statistically significant (r=0.34, P=0.0001). In stage III patients, and in node positive patients, the incidence of hyperprolactinaemia was significantly higher compared to their respective counterparts (stage II vs stage III, r=0.37, P=0.00006; node negative vs node positive, r=0.30, P=0.001). Hyperprolactinaemic patients had a significantly higher risk of developing recurrent/metastatic disease and a higher mortality risk as compared to patients with PRL <20.0 ng/ml. The multivariate survival analysis indicated that apart from disease stage, prognosis of patients with pre-operative hyperprolactinaemia was poorer than that of patients with PRL <20.0 ng/ml. Seventy-eight per cent (87/111) of the tumours showed positive immunoreactivity with PRL antibody indicating that PRL, or a similar molecule, is produced ectopically by breast tumours. PRL mRNA expression using RT-PCR confirmed the de novo synthesis of PRL. PRL mRNA expression was seen in 52% (33/63) of tumours. Sequence analysis of the 234 bp PRL amplimer revealed that the sequence was homologous to the sequence of exon 5 of human pituitary PRL mRNA. Furthermore, PRLR were present in 80% of tumours detected by immunohistochemical localization. A significant positive correlation was noted between IHL-PRL and PRLR (r=0.26, P=0.006). CONCLUSIONS: This multifaceted study of PRL suggests that breast cancer cells produce PRL and that this ectopically produced PRL may act as a major local growth promoter via autocrine and paracrine mechanisms. It may provide new insights into endocrine treatment of breast cancer.

Prognostic value of insulin-like growth factor-1 receptors in patients with colon/rectal cancer: correlation with plasma prolactin.

Prognostic value of IGF-1 receptors (IGF-1R) was evaluated and compared with circulating prolactin (PRL) in 59 patients with Dukes B or C colon/rectal cancer. IGF-1R was estimated by radioligand binding assay and PRL was estimated by immunoradiometric assay. Eighty-five percent (50/59) of patients had IGF-1R- tumours while IGF-1R positivity was observed in only 15% (9/59) of patients. None of the clinicopathological parameters showed any association with IGF-1R status. No significant difference was observed in overall survival period between patients with IGF-1R+ tumours and those with IGF-1R- tumours. However, a significant difference in overall survival time was observed between patients with PRL < 20.0 and > 20.0 ng/ml plasma (X2 = 4.70, df = 1, P < 0.05). In bivariate analysis, patients with IGF-1R- tumours and concomitant hyperprolactinemia had unfavourable prognosis compared to their counterpart (X2 = 4.21, df = 1, P < 0.05). We conclude that there was a trend of better overall survival in patients with IGF-1R+ tumours, and PRL < 20.0 ng/ml plasma when compared to patients with IGF-1R- tumours, and PRL > 20.0 ng/ml plasma. Further, IGF-1R negativity in conjunction with hyperprolactinemia could be used as an indicator of unfavourable prognosis in patients with Dukes B or C colon/rectal cancer.

Can plasma prolactin predict tamoxifen resistance in patients with advanced breast cancer?

The antiestrogen tamoxifen (TAM) is an effective therapy for advanced breast cancer. However, its use is limited by the eventual development of acquired TAM resistance in many patients. There is now strong evidence to suggest that prolactin plays an important role in advanced breast cancer. We have measured plasma prolactin (PRL) and estrogen-receptor (ER) and progesterone-receptor (PR) in post-menopausal patients with breast cancer (Stage III, n = 44). The blood samples were collected pre-operatively and sequentially thereafter for a minimum period of 3 years or until the death of the patients. The ER and PR were estimated in breast tumor samples by dextran-coated charcoal (DCC) method. The patients were treated with surgery and radiotherapy followed by TAM (10 mg, 1 BD). Based on the response to treatments, the patients were divided into two groups: (1) TAM sensitive (n = 19) and (2) TAM resistant (n = 25). In the TAM sensitive group, patients responded to the treatment and did not develop progressive disease within a period of 3 years. On the contrary, in the group of TAM resistant, patients developed progressive disease within a period of 3 years. The development of progressive disease clearly indicated TAM resistance. Plasma PRL correlated well with the disease progression. This study clearly demonstrated that plasma prolactin accurately indicated the response and development of resistance to TAM in patients with advanced breast cancer.

Prognostic significance of plasma prolactin in breast cancer: comparison with the expression of c erb B-2 oncoprotein.

Having established prolactin to be an indicator of disease progression and hyperprolactinemia as an independent predictor of short term prognostication, we have in this report compared plasma prolactin with the expression of c erb B-2 oncoprotein, ER and PR. c erb B-2 oncoprotein, ER and PR are determinants of breast cancer biology. This is a retrospective study of 47 breast cancer patients. When patients were grouped according to the stage of the disease, plasma prolactin was higher in patients with advanced disease than those with stage II disease. The patients were sub-grouped according to prolactin < 20.0 ng/ml and > 20.0 ng/ml. The expression of c erb B-2, ER and PR did not differ in these two sub-groups. The overall survival differed significantly between the two sub-groups of prolactin. The patients were sub-grouped according to c erb B-2 positivity or negativity, there was no significant difference in survival. c erb B-2 expression did not differ between the three grades of the tumor, nodal and receptor positivity or negativity. Hence, the present study reinforces the positive association between hyperprolactinemia and unfavourable prognosis and finds c erb B-2 expression as a weak prognosticator in advanced breast cancer patients.

Evaluation of telomerase activation in head and neck cancer.

During replication of the linear chromosomes, telomeres, i.e. the ends of the chromosomes, are not replicated completely by the conventional DNA polymerases. Therefore, normal somatic cells senesce after certain number of cell divisions. Telomerase is a special reverse transcriptase used by most eukaryotes to achieve immortalization. Telomerase activity has been determined in a variety of cancers. However, there are few reports on telomerase activity in head and neck cancer. The etiology of the disease in India is completely different from Western countries. Tobacco consumption is more prevalent in India and the mode of tobacco consumption (e.g. chewing, snuffing, bidi smoking, reverse smoking) is also different. The present study determined telomerase activity in 32 malignant tumour samples of head and neck cancer patients, 11 samples from patients with precancerous/benign lesions and 30 samples of adjacent normal tissues. Telomerase was found to be activated in 80% of the patients with head and neck cancer, 100% of the patients with precancerous/benign lesions and 74% of the adjacent normal tissues. According to the theory of field cancerization, carcinogenic insults (e.g. tobacco) may result into multiple malignant foci. This fact may explain the reason for high telomerase positivity in adjacent normal as well as precancerous/benign tissues. Telomerase activation and the clinical or histopathological characteristics of the head and neck cancer patients were observed to be independent features. This is a preliminary report which has generated a greater interest for in-depth elucidation of the role of telomerase and telomeres in head and neck carcinogenesis in India.

Circulating prolactin and TPS in monitoring the clinical course of male patients with metastatic tongue cancer: a preliminary study.

Serum prolactin and tissue polypeptide specific antigen (TPS) were assayed pretherapeutically and sequentially thereafter in 20 male patients with advanced (stages III and IV) tongue cancer. The markers were correlated with disease stage, histologic grade and an attempt was also made to compare the predictive value of these markers and two year survival. Prolactin and TPS levels were significantly higher at diagnosis when compared to controls (a = 0.005). Prolactin and TPS levels did not correlate with stage and histologic grade of the tumor. We found that the positive predictive value for prolactin and TPS was 100% and 75% respectively. The two year survival of patients with prolactin > 15.0 ng/ml was 7.88 +/- 1.48 months, compared to 16.98 +/- 1.47 months of those with prolactin less than 15.0 ng/ml (P < 0.001), whereas such a trend was not observed for TPS. This analysis showed an excellent correlation between serial serum prolactin changes and the response to treatment or progression of disease in patients with advanced tongue carcinoma.

Squamous cell carcinoma antigen and protein-bound sialic acid in the management of head and neck cancer.

Serum squamous cell carcinoma antigen (SCCAg) and protein-bound sialic acid (PBSA) were measured in 43 head and neck cancer patients and 50 controls. SCCAg and PBSA were correlated with clinical stage, histological grade, presence/absence of keratin and disease course. Patients with advanced cancer (stage III and IV) and grade III tumors had higher PBSA levels but no such difference was observed for SCCAg. Head and neck cancer patients were grouped according to the disease status i.e. a) patients who developed recurrence and b) who responded to the adjuvant therapies. There was an excellent correlation between serial serum PBSA changes and the progression of disease or the response to therapy in patients with advanced head and neck cancer.

Usefulness of terminal deoxynucleotidyl transferase as prognosticator in leukemia patients.

Peripheral blood samples from 55 previously untreated leukemia patients (33 males, 22 females) were analysed for terminal deoxynucleotidyl transferase (TdT) activity. TdT was significantly higher in patients with acute myelogenous leukemia (AML; P < 0.001), chronic myelogenous leukemia (CML; P < 0.05) and acute lymphoblastic leukemia (ALL; P < 0.001) when compared with controls. One patient with chronic lymphocytic leukemia (CLL) had undetectable TdT. Among leukemic patients, ALL patients had higher concentration of TdT than CML and AML patients. Females had higher TdT activity than males, although the difference between the two groups was not statistically significant. 68% TdT+ and 32% TdT- patients were in blastic crisis. Patients with more than 10% of blasts in the circulation had significantly higher TdT than blast-negative patients (P < 0.001). No difference in survival was observed between TdT+ and TdT- group. From these results, we conclude that the absolute TdT concentration is of little prognostic value in leukemia patients.

Comparison of prolactin, CA 15-3 and TPA in breast carcinomas.

Circulating prolactin, CA 15-3 and TPA were assayed pre-therapeutically and sequentially thereafter from 68 breast cancer patients attending the Gujarat Cancer and Research Institute, Ahmedabad--a regional cancer institute in Western India. The three marker values were correlated with the stage, histologic grade and disease status. At least one of the markers was elevated in 82% of patients. CA 15-3 and TPA levels were elevated with the advancement of stage. Prolactin levels were high in poorly differentiated tumors of pre-menopausal patients. The disease status was effectively reflected by the levels of prolactin and CA 15-3. TPA showed high false positivity so was of no use as an indicator of disease status. Recurrence could be predicted early, with a lead time of 3-6 months using prolactin and CA 15-3.

Role of pretherapeutic biomarkers in predicting postoperative radiotherapy response in patients with advanced squamous cell carcinoma.

PURPOSE: To assess the role of biomarker levels in predicting radiotherapy (RT) response in patients with squamous cell carcinoma of buccal mucosa treated with postoperative RT. MATERIALS AND METHODS: Thirty-one patients with squamous cell carcinoma of buccal mucosa who received postoperative RT were enrolled for the study. Glutathione S-transferase (GST), glutathione reductase (GR), superoxide dismutase (SOD) and catalase activity were analysed from primary tumour and adjacent normal mucosa of the same patients before RT. p53 and p21ras were localized immunohistochemically. RESULTS: Enzyme activation was predicted by comparing the levels of these enzymes in tumour and adjacent normal mucosa. Deactivation of GST, activation of GR, SOD and catalase were associated with poor response to RT. p53 immunoreactivity was associated with failure to respond to RT. CONCLUSIONS: These markers may be useful in predicting treatment outcome in patients receiving postoperative RT, although this conclusion requires confirmation in a larger group of patients.

Mitomycin C induced chromosomal aberrations in young cancer patients.

Mitomycin-C (MMC) induced Chromosomal aberration (CA) frequencies were studied in 48 h peripheral blood lymphocyte (PBL) cultures of untreated cancer patients of young age (maximum age 12 years, n=77). Control population (n=71) consisted of age-matched group (maximum age 12 years, n=21); elder controls (minimum age 60 years, n=19) and healthy first degree relatives, i.e., parents or siblings of the pediatric cancer patients (mean age 24.3 years, n=31) as they share their genome and environment. Induced CA levels were found to be significantly higher among pediatric cancer patients as compared to control groups. The age-matched and elder control groups showed comparable CA levels. The first degree relatives controls showed higher induced CA levels as compared to pediatric and elder control groups. The present results indicate that there are different degrees of mutagen sensitivity prevailing in normal population. This may be responsible for differential cancer proneness. High degree of mutagen sensitivity in cancer patients may also be playing a major role in cancer onset at an early age.

Serum sialic acid forms as markers for head and neck malignancies.

The value of protein-bound (PSA), lipid-bound (LSA) and free sialic acid (FSA) levels in the serum as marker was assessed together with its use as a prognostic indicator for head and neck cancers in 165 patients followed-up for over 18 months. Elevated PSA levels were found in 57% of patients with benign disorders, 52% with primary head and neck cancers, and 75% with metastatic cancer. Strong correlation was found between PSA and the regression/progression of the disease. The PSA level essentially returned to normal in patients with favorable prognosis and PSA levels showed a tendency to increase or remain at high levels in patients with a poor prognosis. These results suggest that PSA level is a better prognostic indicator in head and neck cancers, for which there is no available tumor marker.

Peptide and steroid hormone levels in pre- and postmenopausal breast carcinoma.

Circulating levels of LH, FSH, prolactin, estradiol, progesterone and testosterone were measured by radioimmunoassay in 15 premenopausal (PR-M) age matched healthy controls, 35 premenopausal breast cancer patients prior to therapy, 20 postmenopausal (PO-M) age matched healthy controls and 68-71 postmenopausal breast cancer patients prior to therapy. The patients had histologically proven breast cancer. In PR-M breast cancer group, the LH and progesterone did not differ significantly whereas prolactin showed marked elevation (p less than 0.001) and estradiol and testosterone showed significant decrease (p less than 0.001). The PO-M breast cancer patients exhibited remarkable increase in the levels of LH, FSH, prolactin and testosterone (p less than 0.001) whereas estradiol and progesterone showed little increase in the levels (p less than 0.2 and less than 0.1, respectively). From the results, it is concluded that prolactin and altered ratio of estrogen and androgen plays a major role in the genesis of breast cancer.

Correlation of steroid receptors with histopathologic characteristics in breast carcinoma.

Estrogen and progesterone receptors were correlated to histologic variables in 288 breast cancer patients from western India. The progesterone receptors (PR) were significantly elevated as compared to estrogen receptors (ER) amongst primary pre- and peri-menopausal breast carcinomas. ER positivity was correlated significantly to nuclear grade. Lymphatic invasion and vascular permeation were not found to be linked to ER, while PR positivity could be linked to lymphatic invasion and inversely to vascular permeation. Higher frequency of ER positivity was observed in lobular carcinomas. PR concentrations of the tumor had better correlations with histologic variables as compared to ER.

Estrogen and progesterone receptor status of primary breast cancer: relationship with the pattern of first metastasis and survival.

Estrogen (ER) and progesterone receptor (PR) contents in primary tumors from 127 advanced breast cancer patients were measured by DCC method. The patients were followed for 2 years and the prognostic value of the receptor levels was evaluated and compared with other tumor and patient characteristics. No relation was found between receptor levels and tumor, first relapse site as well as short-term survival (2 years) which might be due to the advanced stage of disease at diagnosis.

Protection from pan masala induced genomic damage by beta-carotene and retinoic acid--an in vitro experience.

Cytogenetic studies in Chinese hamster ovary (CHO) cells using aqueous and organic extracts of pan masalas, as well as genomic damage observed among pan masala consumers have conclusively shown genotoxic potential of pan masala-a dry complex mixture of areca nut, lime, catechu, cardamom, unspecified flavoring agent, etc., often containing tobacco in it. Tobacco and areca nut, major ingredients of pan masala, are closely associated with oral cancer. The most widely studied group of compounds in the field of chemoprevention is retinoids which includes natural vitamin A, beta-carotene and synthetic derivatives of vitamin A. In the present study, antigenotoxic effect of beta-carotene (BC) and retinoic acid (RA) on genotoxic potential of pan masala have been evaluated in CHO cells with the help of sister chromatid exchange (SCE) frequency and chromosome aberration (CA) frequency as cytogenetic markers. The pulse treatment with pan masala plain/pan masala-tobacco (PM/PMT) extract in combination with either BC or RA yielded lower frequencies of CA and SCE in CHO cells as compared to the cultures treated with aqueous extract fo pan masalas alone. This antigenotoxic effect of BC and RA was more pronounced when treatment was given continuously for a longer duration. Thus, these results indicated possibility of using BC and RA to decrease the risk of oral cancer among pan masala chewers.