Recombinant Maspin Protein Inhibits the Tumor Angiogenesis by Regulating SIRT3-Mediated Glycolysis in Cervical Cancer / 中国药学杂志

OBJECTIVE: To investigate the effect of recombinant mammary serine protease inhibitor (Maspin) on tumor angiogenesis and tumor metabolism of HeLa cervical cancer and its mechanisms. METHODS: MTS was used to test the effect of recombinant Maspin on proliferation of HeLa cells. Western blotting was used to determine the effect of recombinant Maspin on the expression of SIRT3 in HeLa cells. SIRT3 siRNA was constructed by commercial and then transfected into HeLa cells. Tube formation assay was used to study the effect of SIRT3 intervene on tumor angiogenesis. Detection kits were used to examine the effects of recombinant Maspin on the glucose consumption and the generation of pyruvic acid, lactic acid as well as ATP after SIRT3 intervene, respectively. Western blotting was used to study the effect of SIRT3 intervene on expression of glucose transporter 1 (GLUT1), monocarboxylate transporter-1 (MCT-1), hexokinase (HK2), fructokinase 6-phosphate(PFK1) and pyruvic acid 2 (PKM2). RESULTS: Compared with the control group, the proliferation of HeLa cervical cancer cells was significant inhibited by recombinant Maspin in vitro. SIRT3 siRNA transfection could significantly decrease the inhibition effect of recombinant Maspin on tumor angiogenesis. SIRT3 siRNA transfection could significantly ameliorate the inhibition effect of recombinant Maspin on glucose consumption and the contents of pyruvic acid, lactic acid as well as ATP, respectively. Recombinant Maspin inhibited the expressions of GLUT1, MCT-1, HK2, PFK1 and PKM2, however, SIRT3 siRNA transfection significantly reversed the expression of above glycolysis-related proteins. CONCLUSION: Recombinant Maspin inhibits the tumor angiogenesis by suppressing glycolysis in HeLa cells, and such effect is dependent on SIRT3 expression.

Expression and significance of Maspin and IKKα in sinusoidal mucosa of rats with fungal rhinosinusitis / 中国比较医学杂志

Objective To investigate the expression and significance of Maspin and IKKα in nasosinusoidal mucosa of rats with fungal rhinosinusitis (FRS).Methods A total of 40 SD rats were used to establish the FRS model, and randomly divided into nasal obstruction group, FRS group, immunosuppressive group and invasive FRS group, 10 rats in each group.Another 10 normal rats were used as control group.Mice in the control group were fed with normal diet.In the nasal obstruction group, the mice had only hemostatic cotton stuffed in the nasal cavity and injection of 0.9% NaCl in the abdominal and nasal cavities.In the FRS group, the mice were injected Aspergillus fumigatus spore suspension into the nasal cavity and 0.9% NaCl i.p.The mice of the immunosuppressive group were given cyclophosphamide i.p.and 0.9% NaCl injection into the nasal cavity.The invasive FRS group was injected with cyclophosphamide i.p.and Aspergillus fumigatus spore suspension into the nasal cavity.The serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).The expression of Maspin and IKKα in nasosinusoidal mucosa was detected by immunohistochemical staining.The expression of Maspin mRNA and IKKα mRNA in the nasosinusoidal mucosa was detected by fluorescence quantitative PCR.Results The serum levels of IL-6 and TNF-α in different groups were significantly different (P 0.05).Theresult of immunohistochemical staining showed that the protein expression of Maspin in the FRS group and invasive FRS group was significantly lower than that in the control group, nasal obstruction group and immunosuppressive group, while the expression of IKKα protein was significantly higher than that of control group, nasal obstruction group and immunosuppressive group (P< 0.05).The protein expression of Maspin in the invasive FRS group was significantly lower than that in the FRS group, by contrast, the expression of IKKα protein was significantly higher (P< 0.05).The PCRresult revealed that the expression levels of Maspin and IKKα mRNA were (0.217 ± 0.013) and (0.193 ± 0.012), significantly lower than that in the control, obstruction and immunosuppressive groups [(0.309 ± 0.021), (0.302 ± 0.017), and (0.293 ± 0.02)] (P< 0.05), while the expressions level of IKKα mRNA were significantly higher [(0.319 ± 0.043), (0.384 ± 0.048) vs (0.169 ± 0.015), (0.171 ± 0.018), and (0.175 ± 0.019)] (P< 0.05).Conclusions Down-regulation of Maspin expression after IKKα activation is the main cause of the onset of FRS, which may also be one of the mechanisms of invasive FRS.

Expression of Maspin in invasive fungal rhinosinusitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore the expression of Maspin in invasive fungal rhinosinusitis (IFRS) and the value of Maspin in the diagnosis of IFRS. METHODS Forty two cases of fungal rhinosinusitis (FRS) were set as the experimental group, which included 12 cases of IFRS and 30 cases of noninvasive fungal rhino-sinusitis (NIFRS). At the same time, 30 cases of chronic rhino-sinusitis were set as control group. Immunohistochemistry (IHC) was used to detect the expression of Maspin. RESULTS Compared with the control group, the expression of Maspin in FRS group decreased statistically (t=-3.367, P<0.05). The IFRS group, compared with other two groups, had the lowest expression of Maspin (t=-3.390, P<0.05; t=-4.143, P<0.05). By using Maspin score of 5.70 as the cut-off point, the sensitivity and specificity for the diagnosis of IFRS was 91.7% and 88.3% respectively. CONCLUSION The expression of Maspin is very low in IFRS group. Down-regulation of Maspin expression may be a potential indicator for diagnosis of IFRS.

Roles of maspin in biological behaviors of A549 cells / 中国病理生理杂志

AIM:To investigate the roles of maspin in the biological behaviors of non-small-cell lung cancer A549 cells.METHODS:Full-length coding region of maspin was amplified by PCR from human normal breast tissue and cloned into MSCV vector .Virus supernatants was produced by Phoenix A packaging system , and target cell line was infec-ted with the virus supernatants .The transfected cells were screened with puromycin .The cell line with maspin over-expres-sion was identified by real-time PCR and Western blot.Cell growth, migration and invasion were investigated by xCelli-gence system .RESULTS:Maspin over-expression vector was successfully constructed .The cell line with maspin over-ex-pression was established .No difference of the growth between A 549-control cells and A549-maspin cells was observed .The migration and invasion were quite different between A 549-control cells and A549-maspin cells.The expression level of inte-grin β1 in A549-maspin cells was decreased compared with A 549-control cells .CONCLUSION:Maspin has no effect on the growth of A549 cells.Maspin suppresses the migration and invasion of A 549 cells, in which integrin β1 is involved.

Expression of maspin in benign and malignant salivary gland tumors: An immunohistochemical study.

Context: Maspin is a novel serine protease inhibitor (serpin) with multifaceted tumor‑suppressive activities. It was originally identified in normal human breast myoepithelial cells and shows variable expression in different types of cancer cells. Maspin displays anti‑metastatic properties in mammary and prostate cancer. Its expression is maintained during ovarian, lung and pancreatic carcinogenesis, indicating that Maspin regulated metastatic potential is tissue specific. Thus, it is possible that Maspin participates in salivary gland tumor biology as well. In this study, expression pattern of maspin in benign and malignant salivary gland tumors is analyzed, to understand the biological behavior of salivary gland tumors with respect to maspin expression. Aims and Objectives: The aim of this study was to demonstrate, record, and correlate the expression pattern of maspin in benign and malignant salivary gland tumors. Settings and Design: A retrospective study of maspin expression in 30 diagnosed cases of benign and malignant salivary gland tumors retrieved from archives of our department. Materials and Methods: Anti‑maspin antibody and horseradish peroxidase detection system. Statistical Analysis: Descriptive statistical analysis and Chi‑square/Fisher Exact test. Results: Intense expression with P < 0.001 is associated with benign tumors, nuclear staining with P < 0.001 is significantly associated with benign tumors and cytoplasmic staining with P = 0.020 is associated with malignant tumors. Conclusion: Intensity of expression is more in benign tumors when compared with malignant tumors. The benign tumors showed both nuclear and cytoplasmic expression. Some malignant tumors did express maspin, but mainly in the cytoplasm.

Maspin gene and the invasion and metastasis of gastric cancer / 国际肿瘤学杂志

The maspin gene,a tumor suppressor gene and a member of the serpin superfamily,is isolated from mammary epithelial cells.Maspin gene plays an important role in the process of invasion and metastasis of gastric cancer.Researches show that maspin gene suppresses the occurrence,development and metastasis of gastric cancer mainly by inducing tumor cell apoptosis,inhibiting tumor angiogenesis,methylation modification,promoting homogeneity adhesion and inhibiting heterogeneous adhesion.

Extracts of Celastrus Orbiculatus Enhance the Invasion Function in Maspin Overexpression Human Gastric Carcinoma MGC803 Cells / 世界科学技术-中医药现代化

This study was aimed to investigate whether the extracts of Celastrus orbiculatus enhanced the invasion function of maspin tumor inhibitor gene through the construction of maspin overexpression human gastric carcinoma MGC803 cell line. Maspin was cloned into plasmid GV208-EGFP eukaryotic expression vector. And then, the recombinant plasmid GV208-maspin-EGFP was transfected into human gastric carcinoma MGC803 cells. After the maspin overexpression MGC803 cell were treated with Celastrus orbiculatus extracts in different concentrations (10, 20, 40 μg·mL-1), the invasion effects were detected by Transwell chamber assay. The results showed that after the successful construction of maspin overexpression cell line, the number of cells invading through Matrigel was obviously decreased in the Transwell chamber assay. It also showed drug concentration dependency. It was concluded that maspin gene can inhibit invasion of gastric carcinoma MGC803 cells. Simultaneously, the extracts of Celastrus orbiculatus can enhance the function of maspin gene.

Maspin gene in tumor / 国际肿瘤学杂志

Maspin,one kind of tumor suppressor gene,belongs to the serine proteinase inhibitor (serpin) ultra family's member.Recent studies found that it suppresses the tumor occurrence and development and has the inhibition of tumor metastasis through suppressing the tumor angiogenesis and increasing the cell adhesion,which may be a new way for gene therapy.

Study on the expression of maspin gene and its role in patients with colorectal cancer / 中华微生物学和免疫学杂志

Objective To investigate the relationship between the expressions of maspin gene in colorectal membrane,and their roles in the occurrence,progression,metastasis and prognosis of colorectal adenocarcinoma.Methods Immunohistochemistry technique was used to detect the expression of maspin in tissues of normal controls,adenoma,colorectal adenocarcinoma primary tumor,liver metastasis,lymph-node metastasis respectively.Results The expressions of maspin in normal controls,adenoma and colorectal adenocarcinoma primary tumor are down-regulated successively; the expression of maspin in colorectal adenocarcinoma with metastasis is lower than that in colorectal adenocarcinoma without metastasis(P＜0.05) ; the down-regulation of maspin expression is significantly correlated with the depth of invasion,differentiation,liver metastasis and lymph node metastasis of colorectal adenocarcinoma(P＜0.01,P＜0.05,P＜0.01,P＜0.05).Conclusion The down-regulation of expression of maspin probably plays an important role in the occurrence,progression and metastasis of colorectal adenocarcinoma.maspin is probable a predictive index for reflecting the metastasis of colorectal adenocarcinoma and provide a clue for early diagnosis of metastasis of colorectal adenocarcinoma.

The Role of Neutrophil Estrogen Receptor Status on Maspin Synthesis via Nitric Oxide Production in Human Breast Cancer / 한국유방암학회지

PURPOSE: Estrogen, through its binding to nuclear estrogen receptor (ER), has been implicated in the development of human breast cancer. The presence or absence of ER in breast lesions has been used to classify breast cancer into ER+ or ER- type. Maspin, an anti-breast cancer protein produced in normal mammary cells, has also been reported to control the condition. Studies have been conducted to determine the role of ER+ and ER- status in neutrophils in the synthesis of maspin in human breast cancer. METHODS: Maspin presence was determined by enzyme linked immunosorbent assay, while nitric oxide (NO) level was determined using the methemoglobin method. RESULTS: Scatchard plots of the equilibrium binding of estrogen demonstrated the presence of 4.18x10(7) receptors per normal neutrophil and 2.46x10(7) receptors per ER+ neutrophil with a similar dissociation constant (0.926 nM). The ER- type showed nonspecific estrogen binding only. At 0.6 nM estrogen, NO synthesis was maximally increased to 1.829 and 0.887 microM NO/10(9) cells at 4 hours in normal and ER+ neutrophils respectively, with synthesis of 2.383 and 1.422 nM maspin in normal and ER+ neutrophils respectively. Estrogen failed to produce these effects in ER- neutrophils. CONCLUSION: ER status in neutrophils determined maspin synthesis in breast cancer through the stimulation of NO synthesis. Neutrophils with ER- status which do not produce any maspin when treated with estrogen, might imply a worse prognostic outcome in ER- breast cancer due to the lack of anti-breast cancer protein synthesis.

Maspin Expression and Its Clinical Significance in Non-Small Cell Lung Cancer / 결핵및호흡기질환

BACKGROUND: Maspin (mammary serine protease inhibitor) is a member of the serpin superfamily. A few studies have examined the role of maspin in tumor suppression of non-small cell lung cancer (NSCLC); however, its role in the development and progression of NSCLC still remains controversial. We evaluated the immunohistochemical expression of maspin in order to elucidate its clinical significance in NSCLC. METHODS: We analyzed 145 patients with pathologically confirmed NSCLC, including 66 cases of squamous cell carcinomas (SCCs) and 79 cases of adenocarcinomas (ADCs). We performed a immuno-histochemical stain with maspin and PCNA (proliferating cell nuclear antigen) using tissue microarray blocks. RESULTS: There were 108 men and 37 women in the study population. The mean age of patients in the study was 63.7 years (range, 40.0~82.0; median, 65.0). The proportion of maspin expression was significantly higher in SCCs (52/66, 78.8%; p<0.01) than in ADCs (17/79, 21.5%; p<0.01). Maspin expression was not associated with PCNA (p=0.828), lymph node involvement (p=0.483), or tumor stage (p=0.216), but showed correlation with well-to-moderate tumor differentiation (p=0.012). There was no observed correlation between maspin expression and survival with NSCLC (p=0.218). CONCLUSION: The present study suggests that maspin expression was significantly higher in SCCs than in ADCs and was associated with low histological grade. However, maspin expression was not an independent factor to predict a prognosis in NSCLC.

The maspin expression in canine mammary tumors: an immunohistochemical and molecular study / A expressão do maspin nos tumores mamários caninos: um estudo imuno-histoquímico e molecular

The serpin maspin, a tumor suppressor in breast cancer was described as an inhibitor of cell migration and inducer of cell adhesion between the basement membrane and extracellular matrix resulting in inhibition of tumor metastasis. In contrast, overexpression of maspin is correlated with poor prognosis in other types of cancer. Little is known about expression, regulation and function of maspin in canine mammary tumors. It was demonstrated in this study, a loss of maspin expression in malignant canine mammary cells compared with a pool of normal canine mammary tissue, analyzed by quantitative real-time PCR; weak maspin expression in malignant canine mammary tumors were observed by immunohistochemistry. It was also demonstrated that a correlation with nuclear maspin expression and a good prognosis. It is suggested that maspin could be used as a prognostic marker in canine mammary neoplasia.

O serpin maspin, um supressor tumoral no câncer de mama foi descrito como inibidor de migração celular e indutor de adesão celular entre a membrana basal e a matriz extracelular resultando na inibição da metástase tumoral. Por outro lado, a alta expressão do maspin está relacionada com um mau prognóstico em outros tipos de câncer. Pouco se sabe sobre a expressão, regulação e função do maspin nos tumores mamários caninos. Neste estudo, foi demonstrada uma perda da expressão de maspin nas células mamárias malignas de cães quando comparadas com um pool de tecido mamário normal de cães, analisado por PCR quantitativa em tempo real. Houve uma expressão fraca maspin em preparações de tumores mamários malignos observadas por imuno-histoquímica. Também foi verificado que a expressão nuclear do maspin em tumores mamários caninos está relacionada a um bom prognóstico. Assim, o maspin pode ser utilizado como um marcador prognóstico nas neoplasias mamárias em cães.

Tumor suppressive properties of maspin and its expression levels in tumors / 肿瘤研究与临床

Maspin (mammary serine protease inhibitor) is a kind of serpin.Many studies have characterized it as a tumor suppressor based on its ability to inhibit tumor angiogenesis,increase adhesion between tumor cells and ECM,inhibit tumor cell invasion and distant metastasis and promote apoptosis.In recent years,some studies have found maspin expresses high in some cancers,such as in mammary cancer and prostate cancer,while in some other cancers,such as in pancreatic cancer,maspin expression level shows high.The expression levels and biological functions of maspin in different organs still need further study.

The expression of cancer suppressor gene maspin in the HCC and its correlation with AFP and its clinical significance / 重庆医科大学学报

Objective:To study the protein expression of maspin in the HCC and its correlation with AFP,and to discuss its clincal significance.Methods:the protein expressions of maspin and AFP in the normal liver tissue(NT),paratumor tissue(PT)and liver tumor tissue(TT)were detected with immunohistoch-emistry and their area densities were measured.A total of30 patients with HCC and 30 patients without HCC in our Department of Hepatobiliary Surgery were as samples from March,2004 to December,2005.Results:(1)The area densities(AD)of maspin protein in the NT,PT and TT were 20 712.48?1 826.28,7 347.40?441.16,85.68?19.81 respectively,but the area densities of AFP protein were 66.00?10.70,7 005.04?335.69,13 094.00?516.32 respectively;the comparion of two was P

Effects of 5-Aza-CdR on Cell Proliferation of Breast Cancer Cell Line MDA-MB-435S and Expression of maspin Gene / 华中科技大学学报（医学）（英德文版）

The effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the proliferation of MDA-MB-435S cells and the expression of tumor suppressor gene maspin were investigated. Human breast cancer cell line MDA-MB-435S was treated with 5 μmol/L 5-Aza-CdR, a specific demethylat-ing agent for 0 to 8 days. The growth of MDA-MB-435S cells was observed by MTT assay before and after 5-Aza-CdR treatment, respectively. The expression of maspin mRNA was detected by re-verse transcription-polymerase chain reaction (RT-PCR). The cell cycle of MDA-MB-435S cells was analyzed by flow cytometry. The results showed that the growth of MDA-MB-435S cells treated with 5-Aza-CdR for 8 days was significantly suppressed as compared with the control groups, and the in- hibition rate increased sharply from 5 day to 8 day (35.42% to 71.29%). Flow cytometry showed that 5 μmol/L 5-Aza-CdR could induce G2/M cell cycle arrest and decrease the percentage of mitosis cell number in this cell line. Maspin mRNA was expressed in MDA-MB-435S cells after 5-Aza-CdR treatment, but it was weakly detectable before the treatment. It was concluded that Maspin gene might be transcriptional silencing by hypermethylation and the re-expression of maspin gene by 5-Aza-CdR can inhibit the proliferation and induce the G2/M arrest of MDA-MB-435S breast cancer cells.

Expression of Maspin is associated with the Intestinal Type of Gastric Adenocarcinoma / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Maspin is known as a tumor suppressor gene, but its significance has been questioned in various human cancers. The aim of this study was to investigate the expression pattern of Maspin in human gastric adenocarcinomas and its possible correlation with clinicopathological findings. MATERIALS AND METHODS: The expression of Maspin mRNA was measured by nested RT-PCR using 60 frozen adenocarcinomas of the stomach and 31 noncancerous tissues from the proximal resection margin. Immunohistochemical study for Maspin protein expression was carried out using 62 paraffin-embedded tissues, composed of both cancer and noncancerous tissues. RESULTS: Maspin mRNA expression was detected in 80.0% (48 of 60) of the gastric adenocarcinomas, but in only 22.6% (7 of 31) of the normal gastric mucosa (p<0.001). The positive rate of Maspin protein expression was higher in the adenocarcinomas than the normal tissues (62.9% vs. 27.4%, p<0.05). In addition, the intestinal type of tumors showed significantly higher expression levels compared to the diffuse type of tumors (81.5% vs. 48.6%, p<0.05). CONCLUSION: Our results suggest that Maspin is frequently expressed in human gastric cancers, and its expression might be associated with tumorigenesis of the intestinal type of gastric cancer.

Prognostic Significance of Maspin in Pancreatic Ductal Adenocarcinoma

BACKGROUND: Maspin is a serpin family of protease inhibitors. Althouth maspin has been considered a tumor suppressor that inhibits the motility, invasion, and metastasis of breast and prostatic cancer cells, there are many conflicting reports about maspin expression and cancer prognosis. METHODS: To investigate whether the expression of maspin could be used as a prognostic marker in pancreatic cancer, 72 paraffin-embedded pancreatic ductal adenocarcinomas were analyzed using immunohistochemistry. We examined the prognostic value of maspin as well as its relationship with clinicopathological features. RESULTS: Maspin expression was observed in all pancreatic ductal adenocarcinoma. Unlike cancer tissues, however, faint or no expression was observed in the corresponding normal pancreatic tissues. In the Cox proportional hazard model, high maspin expression predicted a high hazard rate. Maspin expression had a positive correlation with tumor stage, but there were also no statistically significant relationships between maspin expression and other clinicopathological features. CONCLUSION: These findings suggest maspin expression to have biological relevance in the progression of pancreatic cancers, with potential use as a prognostic marker for pancreatic neoplasm with epithelial origin.

Construction and expression of maspin/pEFIRES-N expression vector / 第三军医大学学报

Objective To construct the expression vector maspin/pEFIRES-N so as to study maspin gene on the inhibition of the growth,invasion,and metastasis in hepatocellular carcinoma.Methods The full lenth of maspin gene was cloned and ligated to the expression vector pEFIRES-N digested by EcoRⅠ+XbaⅠ by T4 DNA ligase.The recombinant vector maspin/pEFIRES-N was stably transfected into hepatocellular carcinoma MHCC-97 cell line,and the expressive changes of maspin gene were detected.Results The recombinant plasmid was amplified in the E.coli.JM109.After the identification and sequencing,the reconstructive plasmid was confirmed containing the correct and full nucleotide sequence of maspin gene and the mRNA and protein level of maspin gene were up-regulated in hepatocellular carcinoma MHCC-97 cells.The proliferation and invasion of MHCC-97 cells was inhibited.Conclusion The expression vector maspin/pEFIRES-N was constructed successfully and could be expressed in eukaryotic cells.

Biological effect of maspin gene on VEGF in hepatocarcinoma cell MHCC-97 / 重庆医科大学学报

Objective:To explore the biological effect of maspin on VEGF(vascular endothelial growth factor,VEGF)in hepatocarcinoma cell MHCC-97 and discuss its significance.Methods:Maspin/pEFIRES-N expressing plasmid was constructed .The changes of maspin and VEGF in the levels of mRNA and protein in hepatocarcinoma cell MHCC-97 before and after the transfection and recombination were detected by RT-PCR and Western blotting.Results:The maspin/pEFIRES-N plasmid was constructed successfully,and was stably transferred into MHCC-97 cell.The expressions of maspin mRNA and protein increased in hepatocarcinoma cell MHCC-97 after the transfection of maspin/pEFIRES-N plasmid,while the expressions of VEGF mRNA and protein decreased ,as compared with those after the transfection of pEFIRES-N plasmid(P

Transfection of maspin/PCR2.1 vector inhibits uPA and uPAR expressions in gastric cancer cells in vitro / 第三军医大学学报

Objective To investigate the inhibitory effect of maspin gene on metastasis and invasion of gastric cancer and its mechanism.Methods Maspin gene was ligated to the expression vector PCR2.1 with T4 DNA ligase after the PCR2.1 was digested by HindⅢ/XbaⅠ.The recombinant vector maspin/PCR2.1 was transfected into human gastric cancer cell lines MKN28 and SGC7901,then the mRNA and protein expression changes of maspin gene,uPA and uPAR were detected respectively by PT-PCR and Western blotting.Results After identified by digestion and sequencing,the reconstructed plasmid was confirmed to contain the correct and full nucleotide sequence of maspin gene.The expressions of maspin gene were up-regulated in MKN28 and SGC7901 cell lines,while the expressions of uPA and uPAR were down-regulated.Conclusion The expression vector maspin/PCR2.1 is constructed successfully and can be expressed in eukaryotic cells.Expressions of uPA and uPAR can be inhibited by maspin gene.