Anti-inflammation treatment for protection of hepatocytes and amelioration of hepatic fibrosis in rats.

Chronic inflammation is considered as an important pathophysiologic mechanism of hepatic cirrhosis, which induces hepatocyte injury and activates hepatic stellate cells (HSCs), thus resulting in hepatic fibrosis. Previous studies have reported that cyclooxygenase-2 (COX-2) inhibitor can effectively treat liver fibrosis, while somatostatin (SST) analogues inhibit the activation of HSCs. The present study aimed to investigate the effects of a COX-2 inhibitor, celecoxib, combined with a SST analogue, octreotide, for protection of hepatocytes and prevention of fibrosis in a rat model of hepatic fibrosis. Therefore, a hepatic fibrosis rat model was established following peritoneal injection of thioacetamide (TAA), and the rats were then treated with a combination of celecoxib and octreotide (TAA + C). Immunohistochemistry and western blotting assays were used to assess the expression levels of proteins associated with inflammation, epithelial-mesenchymal transition (EMT), proliferation, apoptosis and autophagy. H&E staining, transmission electron microscopy and scanning electron microscopy were used to evaluate the destruction of hepatocytes. Masson's Trichrome and Sirius Red were used to measure the degree of liver fibrosis. The results demonstrated that, compared with those of the control group, the degree of liver fibrosis and the expression of the intrahepatic inflammation factors were aggravated in the TAA group. Furthermore, the apoptosis rate, EMT and autophagy of hepatocytes were also increased in the TAA group. However, treatment with TAA + C restored the aforementioned increased levels compared with the TAA group. In conclusion, treatment of rats with the combination of celecoxib and octreotide could attenuate the progress of hepatic fibrosis via protection of hepatocytes by reducing apoptosis, EMT and autophagy in hepatocytes.

Modification of the method to establish a hepatic VX2 carcinoma model in rabbits.

The hepatic VX2 carcinoma model in rabbits is widely used for the preclinical study of hepatocellular carcinoma. In the present study, a modification was made to the conventional method to establish the animal model, as the conventional method gives rise to frequent tumor seeding due to the drop-out of tumor fragments. In order to evaluate each distinct method of establishing the model, the rabbits were divided into two groups: Group A (the conventional method; n=20) and group B (the modified method; n=20). All surgical details were recorded for reference. At 14 days post-surgery, contrast-enhanced computed tomography (CECT) and autopsy were conducted. Microscopic morphology of tumor cells was observed using hematoxylin and eosin (H&E) and transmission electron microscopy (TEM). Vascular endothelial growth factor (VEGF) and cluster of differentiation (CD)31 were detected via immunochemistry and reverse transcription-polymerase chain reaction. In total, 19 rabbits in each group succeeded in model establishment. Throughout the surgery, group A experienced a longer surgery time compared with group B (group A vs. group B, 22.57±1.34 vs. 20.17±1.50 min; P<0.001), an increased tumor fragment drop-out frequency (group A vs. group B, 1.84±0.96 vs. 1.16±0.38; P=0.008) and an increased peritoneal nodule incidence (group A vs. group B, 35 vs. 5%, P=0.042). As for CECT, H&E and TEM, hepatic VX2 allografts in the two groups demonstrated similar imaging presentations and tumor cell morphology. In addition, VEGF and CD31 levels did not differ between the two groups. In conclusion, the modified method for the establishment of hepatic VX2 carcinoma model in rabbits may decrease tumor fragment drop-out frequency during surgery and incidence of tumor seeding without affecting the properties of VX2 carcinoma.

Morphological and molecular variations induce mitochondrial dysfunction as a possible underlying mechanism of athletic amenorrhea.

Female athletes may experience difficulties in achieving pregnancy due to athletic amenorrhea (AA); however, the underlying mechanisms of AA remain unknown. The present study focuses on the mitochondrial alteration and its function in detecting the possible mechanism of AA. An AA rat model was established by excessive swimming. Hematoxylin and eosin staining, and transmission electron microscopic methods were performed to evaluate the morphological changes of the ovary, immunohistochemical examinations and radioimmunoassays were used to detect the reproductive hormones and corresponding receptors. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to test the mtDNA copy number. PCR and western blot analysis were used to test the expression of ND2. The change of morphological features of the rat ovaries revealed evident abnormalities. Particularly, the features of the mitochondria were markedly altered. In addition, reproductive hormones in the serum and tissues of AA rats were also detected to evaluate the function of the ovaries, and the levels of these hormones were significantly decreased. Furthermore, the mitochondrial DNA copy number (mtDNA) and expression of NADH dehydrogenase subunit 2 (ND2) were quantitated by qPCR or western blot analysis. Accordingly, the mtDNA copy number and expression of ND2 expression were markedly reduced in the AA rats. In conclusion, mitochondrial dysfunction in AA may affect the cellular energy supply and, therefore, result in dysfunction of the ovary. Thus, mitochondrial dysfunction may be considered as a possible underlying mechanism for the occurrence of AA.

Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.

Curcumin is potentially therapeutic for malignant diseases. The mechanisms of this effect might involve a combination of antioxidant, immunomodulatory, proapoptotic, and antiangiogenic activities. However, the exact mechanisms are not fully understood. In the present study, we provided evidences that curcumin suppressed the expression of enhancer of zeste homolog 2 (EZH2) in lung cancer cells both transcriptionally and post-transcriptionally. Curcumin inhibited the expression of EZH2 through microRNA (miR)-let 7c and miR-101. Curcumin decreased the expression of NOTCH1 through the inhibition of EZH2. There was a reciprocal regulation between EZH2 and NOTCH1 in lung cancer cells. These observations suggest that curcumin inhibits lung cancer growth and metastasis at least partly through the inhibition of EZH2 and NOTCH1.

RNA sequencing reveals retinal transcriptome changes in STZ-induced diabetic rats.

The present study aimed to investigate changes in retinal gene expression in streptozotocin (STZ)­induced diabetic rats using next­generation sequencing, utilize transcriptome signatures to investigate the molecular mechanisms of diabetic retinopathy (DR), and identify novel strategies for the treatment of DR. Diabetes was chemically induced in 10­week­old male Sprague­Dawley rats using STZ. Flash­electroretinography (F­ERG) was performed to evaluate the visual function of the rats. The retinas of the rats were removed to perform high throughput RNA sequence (RNA­seq) analysis. The a­wave, b­wave, oscillatory potential 1 (OP1), OP2 and ∑OP amplitudes were significantly reduced in the diabetic group, compared with those of the control group (P<0.05). Furthermore, the implicit b­wave duration 16 weeks post­STZ induction were significantly longer in the diabetic rats, compared with the control rats (P<0.001). A total of 868 genes were identified, of which 565 were upregulated and 303 were downregulated. Among the differentially expressed genes (DEGs), 94 apoptotic genes and apoptosis regulatory genes, and 19 inflammatory genes were detected. The results of the KEGG pathway significant enrichment analysis revealed enrichment in cell adhesion molecules, complement and coagulation cascades, and antigen processing and presentation. Diabetes alters several transcripts in the retina, and RNA­seq provides novel insights into the molecular mechanisms underlying DR.

Evidence for transcriptional interference in a dual-luciferase reporter system.

The dual-luciferase reporter assay is widely used for microRNA target identification and the functional validation of predicted targets. To determine whether curcumin regulates expression of the histone methyltransferase enhancer of zeste homolog 2 (EZH2) by targeting its 3'untranslated region (3'UTR), two luciferase reporter systems containing exactly the same sequence of the EZH2 3'UTR were used to perform dual-luciferase reporter assays. Surprisingly, there were certain discrepancies between the luciferase activities derived from these two reporter constructs. We normalized luciferase activity to an internal control to determine the amount of the reporter construct successfully transfected into cells, induced a transcriptional block with flavopiridol, quantified renilla luciferase mRNA levels, and compared the absolute luciferase activity among the different groups. The results suggested that curcumin promoted the transcription of the luciferase genes located downstream of the simian vacuolating virus 40 (SV40) early enhancer/promoter, but not those located downstream of the human cytomegalovirus (CMV) immediate-early or the herpes simplex virus thymidine kinase (HSV-TK) promoters. These results explain the discrepancies between the two luciferase reporter systems. The current study underscores the importance of taking caution when interpreting the results of dual-luciferase reporter assays and provides strategies to overcome the potential pitfall accompanying dual-luciferase reporter systems.

The therapeutic impact of HNP-1 in condyloma acuminatum.

BACKGROUND: Condyloma acuminatum is one of the most commonly occurring sexually transmitted diseases. HNP1 is a small antimicrobial peptide that has been reported to have antiviral activities. AIM: Using the condyloma acuminatum tissue culture to resemble the situation more closely in vivo, we investigate the therapeutic effect of a recombinant plasmid encoding HNP1 gene in condyloma acuminatum tissue. METHODS: Recombinant plasmid DNA carrying HNP1 cDNA was constructed and identified. Then the recombinant plasmid was transfected into a condyloma acuminatum tissue fragment, and the HNP1 expression was determined on these tissue fragments by immunohistochemistry. TUNEL staining and flow cytometry techniques were used to examine cell apoptosis of condyloma acuminatum tissue. Relative real-time polymerase chain reaction was used to validate antihuman papillomavirus therapeutics of the treatment groups. RESULTS: Transfected HNP1 gene was expressed mainly in the cytoplasmic granules of the condyloma acuminatum tissues. Positive apoptotic cells were observed in condyloma acuminatum tissues transfected with the HNP1 gene. In addition, the HPV expression was lower in the HNP1 treatment tissues as compared to their corresponding control tissues. CONCLUSION: The results indicate that HNP1 can directly promote condyloma acuminatum cell apoptosis and play an antivirus role in the condyloma acuminatum tissue by limiting viral replication. These observations suggest a possible application for human HNP1 on condyloma acuminatum therapy.

Reduced BMP6 expression by DNA methylation contributes to EMT and drug resistance in breast cancer cells.

Bone morphogenetic protein 6 (BMP6) is an important regulator of cell growth, differentiation and apoptosis in various types of tumor. In breast cancer, it was considered as a tumor suppressor. Our previous study also confirmed that BMP6 was a critical regulator of breast cancer drug resistance. However, little is known about how its expression is regulated and its mechanisms in breast cancer drug resistance. In the present study, we assessed the DNA methylation regulation of BMP6 based on the presence of a large CpG island in the BMP6 gene promoter. Quantitative DNA methylation analyses showed a significantly increased DNA methylation level in the drug-resistant cell line MCF-7/ADR compared to their parental cells MCF-7. Moreover, the drug-resistant cell line MCF-7/ADR showed an EMT phenotype confirmed by morphology and the expression of EMT marker gene. MCF-7 cells transfected with BMP6-specific shRNA vector also showed an EMT phenotype. The MCF-7/ADR cells treated with the recombinant BMP6 proteins reversed their EMT phenotype. These data indicated that hypermethylation modifications contributed to the regulation of BMP6 and induced an EMT phenotype of breast cancer during the acquisition of drug resistance.

[Expression and clinical significance of bone morphogenetic protein 6 in human breast cancer].

OBJECTIVE: To investigate the expression pattern and clinical significance of bone morphogenetic protein 6 (BMP6) in breast tissues. METHODS: The tumor and adjacent noncancerous tissues were harvested from 36 cases of breast cancer, the expression level of BMP6 mRNA of each sample was measured by quantitative RT-PCR. Immunohistochemistry study was used to examine BMP6 protein expression in 80 cases of breast cancer, then the relationship between the expression of BMP6 and relevant clinical and pathological parameters was analyzed. RESULTS: BMP6 mRNA expression in breast cancer was significantly reduced when compared with normal breast tissues (P< 0.01), BMP6 mRNA level in estrogen receptor-positive (ER) breast cancer was distinctly higher than that in ER breast cancer. The expression of BMP6 mRNA was correlated to tumor grade (P < 0.01). The expression level of BMP6 protein in breast cancer was associated to ER and PR status, histological grade and Ki-67 status (P < 0.05), but not correlated to age, tumor size, human epidermal factor receptor 2 (Her2) status and molecular subtypes of breast cancer (P > 0.05). CONCLUSION: The ectopic expression of BMP6 may play an important role in the development and progression of breast cancer.

Downregulation of BMP6 enhances cell proliferation and chemoresistance via activation of the ERK signaling pathway in breast cancer.

Previous studies indicate that bone morphogenetic protein (BMP) 6 is involved in breast cancer development and progression. However, the mechanism underlying the role of BMP6 in breast cancer cell proliferation, differentiation and chemoresistance remains unknown. In this study, we confirmed that BMP6 expression was downregulated in breast cancer tissues compared with the adjacent normal breast tissues. We further demonstrated that the downregulation of BMP6 was correlated with the estrogen receptor (ER) and progesterone receptor (PR) status, tumor grade and enhanced proliferation (Ki67 proliferation index). In vitro functional experiments showed that the suppression of BMP6 expression by a specific small hairpin (sh)RNA vector led to increased proliferation in the MCF7 breast cancer cell line. Furthermore, knockdown of BMP6 in MCF7 cells enhanced the chemoresistance to doxorubicin by upregulation of mdr-1/P-gp expression and activation of the ERK signaling pathway. Taken together, our data suggest that BMP6 plays a critical role in breast cancer cell aberrant proliferation and chemoresistance and may serve as a novel diagnostic biomarker or therapeutic target for breast cancer.

Effects of exercise intensity on copy number and mutations of mitochondrial DNA in gastrocnemus muscles in mice.

The present study aimed to investigate the effects of exercise intensity on mitochondrial DNA (mtDNA) alterations of copy numbers and mutations in the gastrocnemii of mice. A total of 50 male mice were randomly divided into 5 groups; control group (K) and groups A-D, which underwent 10,30,60 and 90 min of swimming per day, respectively. Samples were obtained after 20 weeks of exercise. Total DNA was collected to analyze mutations in the mtDNA displacement loop (D-loop) regions. mtDNA content was quantified by real­time PCR. Point mutations, monobase insertions and deletions were observed in the mtDNA D-loop regions in the skeletal muscles of the mice. A deletion of 16,232 bp was found in certain groups. The mutation base of the control group was higher than that of the exercise groups. The exercise groups demonstrated significantly altered copy numbers; the 30 min exercise group had the highest copy number. These results suggest that moderate exercise intensity reduces mutations in the mtDNA D-loop regions, and enhances the copy number of mtDNA in the gastrocnemus muscles of mice.

Decreased expression of repulsive guidance molecule member A by DNA methylation in colorectal cancer is related to tumor progression.

Previous studies have shown decreased expression of repulsive guidance molecule member A (RGMa) in colorectal cancer. However, the relationship between the expression levels and promoter DNA methylation status of RGMa and the clinical characteristics of colorectal cancer has not been previously reported. Here, we investigated the expression of RGMa by immunohistochemistry, real-time PCR and western blotting and analyzed the methylation status of the RGMa promoter using Sequenom's MassARRAY platform in colorectal cancer tissues and adjacent normal colorectal tissues. The results showed that RGMa expression was decreased in cancer tissues compared with adjacent normal tissues (p<0.01). Furthermore, a tendency for decreased expression in tumor tissues was observed from Dukes' stage A to stage D (p<0.01). In addition, significantly higher levels of hypermethylation in promoter regions of RGMa were observed in colorectal cancer tissues, compared with those in adjacent normal colorectal tissues (p<0.01). Moreover, the methylation levels of RGMa in tumor tissues were significantly increased in Dukes' stage C and D compared with Dukes' stage A and B (p<0.01). Our results indicate that RGMa expression and promoter methylation status are closely related to colorectal cancer genesis and progression. Determination of the expression level and methylation frequency of RGMa in colorectal cancer tissues may have benefit for early diagnosis and for evaluating patient prognosis.

Systematic analysis of microRNA involved in resistance of the MCF-7 human breast cancer cell to doxorubicin.

Multidrug resistance remains a major clinical obstacle to successful treatment of breast cancer and leads to poor prognosis for the patients. Recently studies have shown that microRNAs play an important role in breast cancer cell resistance to chemotherapeutic agents. In this study, microRNA expression profiles of MCF-7/AdrVp and MCF-7 were analyzed using microarray and the results were confirmed by real-time RT-polymerase chain reaction. Gene Ontology (GO) and pathways mapping tools were employed to analyse systemically the biological processes and signaling pathways affected by differential expression microRNAs. Here, we showed that 181 human microRNAs were differentially expressed between two cell lines. Compared to MCF-7 cells, there were 16 microRNAs down-regulated and 165 microRNAs up-regulated in MCF-7/AdrVp. Western blot confirmed the correlation between specific microRNA expression and corresponding changes in protein levels of their targets, specifically those that have a documented role in cancer drug resistance. Furthermore, we validated that signaling pathway highlighted in the study was involved in drug resistance. These results indicated that breast cancer cell resistant to chemotherapy was associated with a group of microRNAs. GO and pathway mapping are valid and effective approach to analyse the function of microRNAs and the results could be a guideline for further investigation.

[Relationship between photosynthetic characteristics and environmental factors in leaves of Pueraria lobata].

OBJECTIVE: To study the relationship between photosynthetic characteristics and environmental factors in leaves of P. lobata. METHOD: Photosynthetic characteristics and environmental factors were measured by using CIRAS-2 portable photosynthesis system. RESULT: The apparent quantum yield in leaves was 0.0173 micromol CO2 x micromol(-1) photon. The dark respiration rate was 2.9333 micromol x m(-2) x s(-1). The light compensation point of photosynthesis was 180 micromol x m(-2) x s(-1). The light saturation point was 1600 micromol x m(-2) x s(-1). The carboxylation efficiency was 0.0338 micromol x m(-2) x s(-1). The light respiration rate was 2.5 micromol x m(-2) x s(-1). The CO2 compensation point was 100 micromol x mol(-1), The CO2 saturation point was 1 600 micromol x mol(-1). CONCLUSION: Photo flux density and air temperature are major environmental factors influencing diumal changes of net photosynthetic rate.

[Study on the role of Wnt/beta-catenin signaling transduction pathway in hepatocellular carcinoma cell line HepG2 and L02 cell line].

AIM: To explore the possible mechanism of action of Wnt/beta-catenin signaling pathway in hepatocarcinogenesis by investigating the different expressions of the main members on this signaling pathway in hepatocellular carcinoma cell line HepG2 and L02 cell line. METHODS: The mRNAs of Wnt1, Wnt4, beta-catenin, cyclinD1 and c-myc genes were amplified by means of semiquantitative reverse transcription polymerase chain reaction (RT-PCR) in normal liver cell line L02 and hepatocellular carcinoma cell line HepG2, respectively. At the same time, the proteins expression of beta-catenin which was the key member in the Wnt/beta-catenin signaling pathway was examined by immunocytochemical method and Western blot technique. RESULTS: In normal liver cell line L02, the mRNAs of Wnt1, Wnt4, cyclin D1 and c-myc genes were not detected except for the gene of beta-catenin. In hepatocellular carcinoma cell line HepG2, the mRNAs of Wnt1, beta-catenin, cyclin D1 and c-myc genes were detected except for the gene of Wnt4. Meanwhile, found that beta-catenin proteins were accumulated in the cytoplasm and/or nucleus in HepG2 but only in cell membrane in L02.Using Western blot technique, found that beta-catenin proteins expression was higher in HepG2 than in L02. CONCLUSION: The Wnt/beta-catenin signaling transduction pathway is activated with aberrant expression of Wnt1 in hepatocellular carcinoma cell line HepG2.

[Separation and determination of puerarin, daidzin and daidzein in stems and leaves of Pueraria thomsonii by RP-HPLC].

OBJECTIVE: To separation and determine the contents of puerarin, daidzin and daidzein in the stems and the leaves of Pueraria thomsonii, and to provide scientific basis for developing and using of the stems and the leaves. METHOD: A RP-HPLC method was applied with a Diamonsil C18 column (4.6 mm x 150 mm, 5 microm) by gradient elution using methanol-1% glacial acetic acid solution as the mobil phase. The flow rate was 1 mL x min(-1) and the detective wavelength was 250 nm, the column temperature was 25 degrees C. RESULT: All of the three compounds showed good linearities (r >0.9995) and the recoveries were in the range of 99.0% - 101.6%. The contents of puerarin, daidzin and daidzein in the stems are higher than those in the leaves. CONCLUSION: The method was accurate and could be used to contral the quality of the stems and leaves of P. thomsonii.

[Expression pattern of nitric oxide synthase in the retina of diabetic rats].

OBJECTIVE: To investigate the expression of nitric oxide synthase (NOS) in the retina of 8-week-old diabetic rats, and explore the potential molecular mechanisms for the role of NO in diabetic retinopathy (DR). METHODS: Retinal gene expression profile of normal and 8-week-old diabetic rats was constructed with restriction fragment differential display polymerase chain reaction (RFDD-PCR). Bioinformatic analysis of the differentially expressed gene identified the genes coding for 3 subtypes of NOS, namely eNOS, nNOS and iNOS as the candidate genes related to DR, which was verified using semi-quantitative RT-PCR and immunohistochemistry. RESULTS: The results of RFDD-PCR revealed down-regulated expression of eNOS and nNOS and up-regulated iNOS expression in diabetic rat retina. RT-PCR showed that the expression levels of eNOS and nNOS in diabetic rat retina were obviously lower than that in normal retina (0.23-/+0.03 vs 0.32-/+0.03 for eNOS, P<0.05; 0.25-/+0.02 vs 0.36-/+0.02 for nNOS, P<0.05), but the expression level of iNOS obviously higher (0.27-/+0.02 vs 0.20-/+0.03, P<0.05). Immunohistochemistry of healthy retina visualized eNOS-, nNOS- and iNOS-positive cells, all located in the inner nuclear layer (INL) and ganglion cell layer (GCL), and eNOS-positive cells were also found in vascular endothelium. In diabetic retina, the number of eNOS- and nNOS-positive cells was significantly lowered in comparison with normal rat retina (14.33-/+3.19 vs 22.13-/+3.60 for eNOS, P<0.05; 21.87-/+3.62 vs 34.40-/+7.09 for nNOS, P<0.05), but the number of iNOS-positive cells significantly increased (17.60-/+2.58 vs 11.73-/+2.70, P<0.05). CONCLUSION: The alterations in eNOS, nNOS and iNOS expression are associated with the deuelopmant and progression of DR.

[Expression of liver-type fatty acid-binding protein and vascular endothelial growth factor and their correlation in human hepatocellular carcinoma].

OBJECTIVE: To detect the expression of liver-type fatty acid-binding protein (L-FABP) and vascular endothelial growth factor (VEGF) in human hepatocellular carcinoma (HCC) and its adjacent liver tissues, and investigate the correlation between the expressions of L-FABP and VEGF and their role in the occurrence and progression of HCC. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical (IHC) staining were employed to examine the expression of L-FABP and VEGF in HCC and its adjacent liver tissues obtained from the surgical specimens of 61 HCC patients who underwent liver resections in West China hospital. RESULTS: The results of RT-PCR showed that the expression level of L-FABP and VEGF in HCC was significantly higher than that in its adjacent liver tissues (L-FABP: 0.97-/+0.12, 0.83-/+0.14, t=5.21, P<0.05; VEGF: 0.92-/+0.11, 0.59-/+0.15, t=11.79, P<0.05). L-FABP tended to co-express with VEGF (P<0.05). IHC staining revealed that the expression of L-FABP and VEGF was mainly located in the cytoplasm, and the gray scale of L-FABP expression was significantly higher than that in the adjacent liver tissues (92.73-/+7.67, 82.83-/+6.90, t=7.44, P<0.05). The number of L-FABP- and VEGF-positive cells in HCC was significantly lower than that in the adjacent liver tissues (L-FABP: 92.18-/+4.44, 84.52-/+6.43, t=5.94, P<0.05; VEGF: 88.69-/+5.56, 77.64-/+5.93, t=8.72, P<0.05). Co-expression of L-FABP and VEGF observed in RT-PCR and also in IHC (P<0.05). CONCLUSION: Both L-FABP and VEGF expressions are up-regulated in HCC. L-FABP gene may be involved in the carcinogenesis of human HCC. Expression of L-FABP is associated with VEGF expression, suggesting that L-FABP promotes the growth of blood vessels by taking up the fatty acids from the bloodstream, and both of them produce a marked effect on energy metabolism in HCC.

[Expression of fatty acid binding protein in human breast cancer tissues].

AIM: To detect the expression pattern of fatty acid binding protein (FABP) in human breast cancer, and to find potential markers and therapeutic targets for breast cancer. METHODS: adipocyte-FABP (A-FABP), heart or muscle FABP (H-FABP), brain-FABP (B-FABP), epidermis or psoriasis FABP (E-FABP), liver FABP (L-FABP), intestinal FABP (I-FABP) and gastro-FABP (G-FABP) expression in 35 ductal infiltrating carcinoma and 16 fibroadenoma of breast were detected by RT-PCR, immunohistochemical staining and Western blot analysis, respectively. RESULTS: E-, L-, and H-FABP were up regulated significantly in ductal infiltrating carcinoma when compared with those in benign tissue (P<0.05). However, there were not significant difference in A-, B-, G-, and I-FABP expression between ductal infiltrating carcinoma and benign tissue (P>0.05). Interestingly, H-FABP was not only found in benign tissues but in some of ductal infiltrating carcinomas, furthermore, H-FABP level was elevated in malignant tissue, compared with that in benign tissue. CONCLUSION: E-, L-, and H-FABP may play a key role in the progress of invasiveness and metastasis in human breast cancer. Furthermore, the secretion of these FABPs has the potential to serve as a diagnostic marker for breast cancer.

Comparative proteome analysis of breast cancer and adjacent normal breast tissues in human.

Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS), incorporated with online database searching, were performed to investigate differential proteins of breast cancer and adjacent normal breast tissues. Considering that serum albumin is abundantly presented in normal control samples, 15 differential spots detected in 11 out of 12 (91.7%) breast cancer samples were identified by online SIENA-2DPAGE database searching and MALDI-TOF/TOF-MS analysis. The results indicate that pathological changes of breast cancer are concerned with augmentation of substance metabolism, promotion of proteolytic activity, decline of activity of some inhibitors of enzymes, and so on. Some important proteins involved in the pathological process of breast cancer with changed expression may be useful biomarkers, such as alpha-1-antitrypsin, EF-1-beta, cathepsin D, TCTP, SMT3A, RPS12, and PSMA1, among which SMT3A, RPS12, and PSMA1 were first reported for breast cancer in this study.