Prognostic significance of FOXA1 and BRCA1 expression in triple negative breast cancer

AIM: To investigate the expression of forkhead box protein A1(FOXA1) BRCA1 protein,P53 and vascular endothelial growth factor (VEGF) in triple negative breast cancer (TNBC) and non-TNBC, and the relevance with the clinicopathological parameters for evaluating the prognosis. METHODS: The tumor samples were collected from 113 cases of breast cancer patients in the First Affiliated Hospital of Jinan University,and divided into TNBC group, luminal subtype group and HER-2 overexpression subtype group by the immunohistochemical results of estrogen receptor, progesterone receptor and HER-2. EnVision two-step method was used to detect the expression of FOXA1, BRCA1, P53 and VEGF in the tumor samples. RESULTS: Total FOXA1 positive expression rate was 63.7% (72/113), with 45.2% (19/42) in TNBC, 88.0% (44/50) in luminal subtype and 42.9% (9/21) in HER-2 overexpression subtype.The statistically sigfnificant difference among the 3 groups was observed (P<0.01). Total BRCA-1 positive expression rate was 47.8% (54/113), with 66.7% (28/42) in TNBC, 44.0% (22/50) in luminal subtype and 19.0% (4/21) in HER-2 overexpression subtype.The statisticallysignificant difference among the 3 groups was also observed (P<0.01). In the cases of clinical stages Ⅰ~Ⅱand histological grades 1~2, FOXA1 positive rate was higher than the FOXA1 negative rate (P<0.01). Negative correlations between FOXA1 positive rate and expression of P53/VEGF, and between FOXA1 positive rate and the recurrence rate were found (P<0.05). In the cases of clinical stages Ⅱ~Ⅲ and histological grades 2~3, the BRCA1 positive rate was higher than the BRCA1 negative rate (P<0.05). Positive correlations between BRCA-1 positive rate and the expression of P53/VEGF, and between BRCA1 positive rate and the recurrence rate were also observed (P<0.05). CONCLUSION: Expression of FOXA1 and BRCA1 in breast cancer is different. BRCA1 may be an adverse prognostic indicator for triple negative breast cancer.     

Construction of siRNA-survivin and GRIM-19 coexpression plasmid and its growth inhibitory effect on prostatic cancer DU145 cells

AIM: To construct the recombinant plasmid that expresses siRNA-survivin and GRIM-19 simultaneously, and to identify the validity of the recombinant plasmid and observe its effect on expression of survivin and GRIM-19 and proliferation ability of prostate cancer DU145 cells. METHODS: The recombinant plasmid coexpressing siRNA-survivin and GRIM-19 was constructed using gene cloning technique. The prostatic cancer DU145 cells were transfected with the coexpression plasmid and control plasmids. Survivin and GRIM-19 mRNA expression was detected by semi-quantitative RT-PCR. The proliferation ability affected by coexpression plasmid was measured by MTT assay. RESULTS: The coexpression plasmid pGRIM-19-si-survivin was successfully constructed according to DNA recombinant technique and identified through restriction enzyme digestion and plasmid sequencing. Compared with the mock, survivin mRNA expression levels were 0.55?0.05,0.62?0.08 and 0.35?0.05 in psi-survivin, pGRIM-19 and pGRIM-19-si-survivin groups, respectively. Compared with psi-survivin and pGRIM-19, pGRIM-19-si-survivin inhibited survivin mRNA expression markedly (P<0.05), while the expression levels of GRIM-19 mRNA were 1.93?0.14, 2.57?0.20 and 4.12?0.21 in psi-survivin, pGRIM-19 and pGRIM-19-si-survivin groups, respectively (P<0.01). Compared with pGRIM-19 group, pGRIM-19-si-survivin enhanced GRIM-19 mRNA expression more obviously (P<0.05). After transfection for 48 h, the proliferation rates were 58.0%?7.2%, 62.1%?6.1% and 50.2%?4.8% in the 3 experiment groups compared with the mock (P<0.05). After transfection for 72 h, the proliferation rate were 43.4%?4.3%, 51.3%?6.7% and 26.8%?7.1% in experiment groups compared with the mock (P<0.05). Compared with psi-survivin and pGRIM-19, pGRIM-19-si-survivin significantly inhibited the cell growth (P<0.05). CONCLUSION: Transfection of coexpression plasmid pGRIM-19-si-survivin dramatically changes the mRNA expression of survivin and GRIM-19 and inhibits the cell growth.     

Expression of DNMT in gastric cancer and effects of DNMT inhibitor 5-aza-2’-deoxycytidine on gastric cancer

AIM: To investigate the protein expression of DNA methyltransferases (DNMT1, DNMT2, DNMT3A, DNMT3B and DNMT3L) in gastric cancer and the effects of DNMT inhibitor (5-aza-2’-deoxycytidine) on gastric cancer. METHODS: The method of immunohistochemistry was used to detect DNMT expression in 60 pairs of gastric cancer and corresponding non-cancerous tissues. The expression of DNMT in gastric cancer cells (SGC-7901 and MKN-45) and gastric mucosa cells (GES-1) was detected by immunofluorescence and Western blotting. The proliferation, cell cycle and apoptosis in the cell lines of SGC-7901, MKN-45 and GES-1 were determined by MTT assay and flow cytometry with 5-aza-2’-deoxycytidine treatment. RESULTS: Both gastric cancer and corresponding non-cancerous tissues expressed 5 kinds of DNMT proteins, but the expression levels of DNMT1 and DNMT3A were significantly lower in gastric cancer than those in non-cancerous tissues (P<0.01). The expression levels of DNMT2 and DNMT3L were also lower in the cancer than those in non-cancerous tissues (P<0.05). No difference of DNMT3B expression level between cancer and non-cancerous tissues was observed (P>0.05). On the other hand, the cells lines of SGC-7901, MKN-45 and GES-1 also expressed DNMT proteins. Except DNMT3B, no difference of the other kinds of DNMT between gastric cancer cells and gastric mucosa cells was observed. DNMT inhibitor 5-aza-2’-deoxycytidine did not have any effect on the growth rate, cell cycle distribution or apoptotic rate in gastric cancer cells and gastric mucosa cells. CONCLUSION: Gastric cancer expresses low levels of DNMT proteins than normal gastric mucosa. 5-Aza-2’-deoxycytidine is not useful for treating gastric cancer.     

Circadian protein Cry2 is a prognostic factor of colorectal cancer and a predictor of chemotherapy

AIM:To assess the association between the expression of Cry2 and the prognosis of colorectal cancer for determining the role of Cry2 in predicting the outcome of chemotherapy. METHODS:Sixteen primary colorectal cancer patients who consecutively underwent neoadjuvant chemotherapy with 5-fluorouracil were enrolled in the present study. The tumor specimens were obtained by colonscopy prior to treatment. The tumor response was evaluated by the response evaluation criteria in solid tumors(RECIST). Accordingly, the patients were divided into 2 groups:complete response(CR)/partial response(PR) and stable disease(SD)/progress disease(PD). Two-tail ² test was applied to analyze the data. In parallel, we assessed 307 patients, who underwent tumor resection between 2001 and 2005. Survival time was calculated from the date of surgery to the date of death or the last follow-up date. Survival curves were obtained by Kaplan-Meier method. Log-rank test was applied for univariate analysis. The Cox proportional hazards regression model was used to identify the independent prognostic factors. RESULTS:The expression of Cry2 was high in CR/PR patients and was low in SD/PD patients, with significant difference(P<0.05). The average survival time of the 84 patients with high expression of Cry2 was 83.458 months, while the average survival time of 223 patients with low expression of Cry2 was 100.10 months, also with significant difference(P<0.05). The patients with low expression of Cry2 lived longer. The multivariate Cox regression analysis indicated that the expression of Cry2 was an independent prognostic factor for colorectal cancer patients(HR was 1.70, 95% CI was 1.09 to 2.66, P<0.05). Multivariate Cox regression showed that high Cry2 expression predicted a worse prognosis(HR was 2.88, 95%CI was 1.03 to 8.06, P<0.05) in colorectal cancer patients with neoadjuvant chemotherapy. CONCLUSION:Expression of Cry2 is an independent prognostic factor for colorectal cancer patients. Lower expression of Cry2 indicates good response to neoadjuvant chemotherapy.     

Alpha basic crystallin expression in canine mammary tumors

The aim of this study was to evaluate prognostic and/or diagnostic factors of canine mammary tumors by immunohistochemically analyzing the expression of alpha basic crystallin (αB-c). For this, formalin-fixed, paraffin-embedded blocks of 51 naturally-occurring canine mammary tumors (11 benign and 40 malignant) were used. Tissue from eight normal canine mammary glands were served as a control. Immunohistochemically, in the control mammary tissues, a few luminal epithelial cells were αB-c positive but myoepithelial cells were negative. In benign or simple type malignant tumors, αB-c expression was observed in luminal epithelial cells while the myoepithelial basal cells were negative. In benign or complex type malign tumors, positive staining was predominantly found in the cytoplasm of epithelial cells. Immunoreactivity of αB-c was also observed in neoplastic myoepithelial cells. Statistically, the number of cells immunolabeled with αB-c was found to be significantly different among tissues from normal canine mammary glands, benign lesions, and malignant tumors (p < 0.05). αB-c immunoreactivity was higher in malignant tumors than the control mammary tissues (p < 0.001). Data obtained in the current study revealed a strong association between high expression levels of αB-c and primary mammary gland tumors in canines.     

Mortality,morbidity, and external injuries in piglets housed in two different housing systems

An investigation was undertaken to study two different types of farrowing pens, use being made of well characterized veterinary parameters for the well‐being of piglets. A housing system with a solid floor and straw‐bedding was compared with a partly metal‐slatted floor system. The veterinary parameters we used were mortality, morbidity, and external injuries in piglets. There were slight differences in mortality and morbidity rates between the two housing systems. There were big differences with regard tp the piglets” injury‐index’ of the two housing systems. The straw system was more favourable. The studies show that obervations of external injuries ('Method of Ekesbo') can be used to judge housing systems of piglets in the farrowing house.     

Acute myelomonocytic leukaemia with short-term spontaneous remission in a cat

A 2-year-old, spayed female domestic shorthair cat was referred with a history of anorexia and depression of 1 week duration. On physical examination, the cat was lethargic and febrile, with splenomegaly, anisocoria and ulcerative stomatitis. A complete blood count (CBC) and a biochemistry profile showed leukocytosis, numerous blast cells in the peripheral blood, thrombocytopenia, hyperglobulinaemia and a positive test for feline leukaemia virus antigen. A diagnosis of acute myelomonocytic leukaemia was made on the basis of the results of bone marrow cytology, histopathology, and immunochemistry (CD3, CD79a, lysozyme, and myeloperoxidase) tests. Following an unexpected 1-month period of clinical and clinicopathological remission without chemotherapy, the cat relapsed and died 1 week later.     

Retrospective – systematic study and quantitative analysis of cellular proliferation and apoptosis in normal, hyperplastic and neoplastic perianal glands in dogs

Neoplasms in the perianal region are frequently diagnosed in dogs. The aetiology is unknown, and most of them are benign. In this study, 240 neoplasms of the perianal glands of dogs were retrieved from the Department of Pathology archives of the Faculty of Veterinary Medicine and Zootechny of University of São Paulo (FMVZ/USP), from 1984 to 2004. All 240 cases were re‐examined by two pathologists. Nine cases (4%) were diagnosed as hyperplasia, 49 (20%) as group I adenoma, 81 (34%) were classified as moderately differentiated adenomas of the group II, 46 (19%) were poorly differentiated adenomas of group II, 48 (20%) were carcinoma of the group III according to the classification proposed by Berrocal, and 7 (13%) were other kind of tumours. Males over 8 years of age were predominantly affected. Cell proliferation was quantified by counting proliferating cell nuclear antigen (PCNA) positive nuclei, and apoptosis was quantified by counting fluorescent eosin‐stained apoptotic corpuscles (AC) in normal tissue, hyperplasia and in different histologic types of neoplasia of these glands. A parallel pattern of increase in both parameters (cell proliferation and apoptosis) was obtained. The net growth index (NGI), represents how much a cell population is proliferating or dying and was achieved by dividing the mean PCNA count in 1000 cells by the mean AC stain count in 1000 cells. NGI was different between hyperplasia and neoplasia; group I adenomas have a much higher potential of growth, and NGI decreases from benign towards malignant lesions. These results show up the importance of studying cell proliferation and apoptosis to understand the carcinogenesis of dog perianal gland.     

《园艺学报》征稿简则

AIM:To investigate the prognostic value of biomarkers on predicting recurrence of hepatocellular carcinoma (HCC) after orthotopic liver transplantation (OLT). METHODS:Fifty patients with HCC who underwent OLT between April 2002 and November 2005 with a minimum clinical follow up of 12 months were included in this retrospective study. We examined immunohistochemical expression of E-cadherin with β-catenin, Ki-67 proliferative index and analyzed the correlation between these biomarkers with recurrence and survival, along with the main clinical-pathological variables.RESULTS:Only at univariate analysis, TNM, portal vein tumor thrombi were valuable on predicting recurrence and survival time (P<0.05), and preoperative serum AFP correlate to recurrence only at multivariate analysis (OR=2.552, P<0.05). Lower membrane expression of E-cadherin, nuclear β-catenin localization and high Ki-67 index showed notable significance on predicting recurrence and survival time at univariate analysis, as well at multivariate analysis, all P<0.01. The membrane expression of β-catenin did not correlate to the prognosis (P>0.05).CONCLUSION:These three biomarkers might have potential as a tumor prognostic marker for predicting recurrence of HCC after OLT and perhaps are better than the clinical-pathological variables.     

DIM enhances effect of cis-diamminedichloroplatinum on growth inhibition and apoptosis in human prostate cancer cell PC-3

AIM: To study the effects of the combination of cis-diamminedichloroplatinum(DDP) and 3, 3-diindolylmethane (DIM) on the growth and apoptosis of human prostate cancer cell PC-3. METHODS: MTT method was applied to detect the cell growth inhibitory rate. The cell apoptosis was measured by the flow cytometry and acridine orange staining method. The expression of the anti-oncogene p21 was detected by RT-PCR technique. RESULTS: The combination of 60 μmol·L^-1 DIM and 0.4 mg·L^-1 DDP effectively inhibited the growth and induced apoptosis in PC-3 cells. This result was the same as the effect of using 4 mg·L^-1 DDP only. The cell growth inhibitory and apoptosis rates for the combination of DIM and DDP were much higher than those for the individual effect. Both the combination and the single effect of these two medicines (i.e., DIM and DDP) all strengthen p21 mRNA expression significantly, and the effect of combination was more significant. CONCLUSION: DIM significantly enhances the effects of DDP on the growth inhibition and apoptosis induction in PC-3 cells.