Expression of calponin-1 and its pathogenic role in systemic sclerosis / 南方医科大学学报

OBJECTIVE@#To investigate the expression of calponin-1 (CNN1) in systemic sclerosis (SSc) and its pathogenic role in fibrosis.@*METHODS@#Skin biopsy samples were collected from 19 patients with SSc and 21 healthy subjects. Real-time PCR was used to detect the expression of and mRNAs in the samples, and the protein expression of CNN1 was detected using immunohistochemistry. In cultured primary human dermal fibroblasts, expression was knocked down RNA interference, and the mRNA expression levels of and the fibrosis-related genes , , , , and were detected using real-time PCR; the proliferation of the cells was assessed using a real-time cell proliferation detection system.@*RESULTS@#Compared with that in samples from normal subjects, the expression of mRNA was significantly increased in the skin tissue of patients with SSc ( < 0.05) with a positive correlation with α-SMA (=0.7219, < 0.0001); the protein expression of CNN1 was also significantly increased in the skin tissue of patients with SSc. In cultured primary skin fibroblasts, the expression of CNN1 mRNA was positively correlated with and mRNA expressions (=0.6547, < 0.05; =0.6438, < 0.05). knockdown in the fibroblasts significantly inhibited the cell proliferation, obviously lowered the expressions of fibrosis-related genes, and reduced the protein expression of collagen.@*CONCLUSIONS@#The expression of is increased in the skin tissues of patients with SSc, and knockdown can reduce the activity of dermal fibroblasts, suggesting the close correlation of CNN1 with fibrosis in SSc.

Calponin Isoforms and Biological Functions / 现代生物医学进展

Calponin is an actin filament-associated regulatory protein expressed in smooth muscle and many types ofnonmuscle cells.Three calponin isoforms 1,2,and 3 are encoded by three homologous genes CNN1,CNN2 and CNN3 in vertebrate species with cell type-specific expression and functions.Besides modulating the activity of smooth muscle myofilaments and contractility,calponin also regulates the functions of actin cytoskeleton in non-muscle cells during cell proliferation,adhesion,migration,differentiation,phagocytosis and fusion.This review focuses on the evolution,tissue and cell type-specific expression,structure-function relationships,transcriptional regulation and relevant mechanisms.

Comparison of estrogen receptor-alpha, progesterone receptor and calponin expression in gonadotrophin-releasing hormone agonist-sensitive and -resistant uterine fibroids

OBJECTIVE: This study was aimed to compare immunohistochemical expression of estrogen receptor (ER)-alpha, progesterone receptor (PR), and calponin in gonadotrophin-releasing hormone agonist (GnRH-a)-sensitive and -resistant uterine fibroids. METHODS: We collected data retrospectively. The sensitive group consisted of women who had reduction in uterine volume greater than 40% following GnRH-a treatment. Uterine volume was either reduced by less than 10%, or was increased in the resistant group. A tissue microarray was constructed using formalin-fixed, paraffin-embedded tissues, 31 and 26 patients for the sensitive and resistant groups, respectively. Tissue sections were immunostained with antibodies against ER-alpha, PR, and calponin. The intensity and area of the immunohistochemical reactions were evaluated using a semi-quantitative scoring system. The Mann-Whitney U-test, Fisher's exact test, and Spearman's rank correlation test were used for analysis of data. RESULTS: PR (P = 0.04) and calponin (P = 0.03) showed a significantly higher staining intensity in the resistant group than in the sensitive group. Both groups showed comparable expression of ER-alpha (P = 0.23). In correlation analysis between changes in uterine volume after GnRH-a therapy and clinicopathological factors, the immunohistochemical intensity of PR (P = 0.04) and calponin (P = 0.03) was significantly correlated with changes in uterine volume. CONCLUSION: This study shows that GnRH-a resistance of uterine fibroids is not related to ER-alpha content, but the expression of PR and calponin is related with GnRH-a resistance.

Expression of calponin in periglomerular myofibroblasts of rat kidney with experimental chronic injuries / 대한해부학회지

Our previous research demonstrated that calponin-immunoreactivity was localized in myofibroblasts of the periglomerular region of human kidney specimens obtained at the time of transplantation from organ recipients. In the present study we examined calponin expression in two chronic nephropathy models, puromycin aminonucleoside (PAN) nephropathy and subtotal nephrectomy (SNx), to investigate the role of calponin in chronic renal injury. Male Sprague-Dawley rats were used, and both nephropathy models were established at 1, 2, 4, and 8 weeks after surgery. There were no periglomerular calponin-positive cells in sham, PAN 1 and 2 week, and SNx 1, 2, and 4 week groups. In SNx 8 week and PAN 4 and 8 week groups, only a few glomeruli with periglomerular calponin-reactivity, which covered half or a very small part of the periglomerular space, were observed. All glomeruli with periglomerular calponin-reactivity showed sclerotic changes, especially thickening of parietal epithelial cells (PECs). In conjunction with our previous report, this data represents the first documentation of the expression of calponin in renal myofibroblasts. We suggest that interactions between PECs and calponin-positive myofibroblasts may play a key role in the late stage of glomerulosclerosis.

Construction of adenoviral vector encoding Calponin-1 siRNA and its effect on human myometrium cells in vitro / 中南大学学报(医学版)

Objective To investigate the effect of Calponin-1 suppression on human myometrium cells through adenovirus mediated siRNA. Methods Human uterine smooth muscle tissues were digested with enzymes, cultured and confirmed with immunocytochemistry. Aadenovirus siRNA-Calponin-1 plasmid was transfected into primary cultured uterine smooth muscle cells in vitro. The expressions of Calponin-1 mRNA and protein were analyzed by RT-PCR and Western blot, respectively.Results The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid was successfully constructed, and Calponin-1 siRNA mediated by recombinant adenovirus resulted in markedly reduced expression of Calponin-1 mRNA and protein in human myometrium cells. The gray values of Calponin-1 mRNA in the uterine smooth muscle cells in the experimental, blank control, and empty vector groups were 316.3±39.2, 1048.5±126.4 and 1027.2±127.5, respectively. The gray values of Calponin-1 protein were 323.3±43.2, 1021.5±143.4, and 1019.2±144.5,respectively. The difference between the experimental group and the blank control group as well as the empty vector group was significant (P< 0.05). There was no significant difference between the empty vector group and the blank control group (P>0.05).Conclusion The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid can inhibit the expression of Calponin-1 in human myometrium cells in vitro,which may be a useful approach to determine the role of Calponin-1 in delivery.

Immunohistochemical expression of vimentin, calponin and HHF-35 in salivary gland tumors

Myoepithelial cells present a complex immunophenotype, with the expression of proteins varying according to the stage of normal or neoplastic differentiation of the cell. In order to evaluate the immunohistochemical markers expressed by these cells, a panel of antibodies composed of vimentin, calponin and HHF-35 was applied to 28 salivary gland tumors. The results demonstrated a higher percent sensitivity of vimentin and calponin compared to HHF-35. However, calponin and HHF-35 presented a focal labeling pattern in contrast with the diffuse distribution of vimentin. The cells predominantly stained by all tested antibodies included nonluminal cells in duct-like and tubular structures, such as those seen in pleomorphic adenomas and adenoid cystic carcinomas, as well as cells in the cords and nests of polymorphous low-grade adenocarcinomas and peripheral cells of sheets and nests of myoepitheliomas. In conclusion, the combination of calponin and vimentin is suggested for the identification of myoepithelial cells in salivary gland tumors.

As células mioepiteliais apresentam um imunofenótipo complexo, variando a expressão de suas proteínas na dependência do seu estágio de diferenciação normal ou neoplásico. Com o objetivo de avaliar comparativamente marcadores imuno-histoquímicos para estas células, um painel de anticorpos composto pela vimentina, calponina e HHF-35 foi aplicado em 28 tumores de glândulas salivares. Os resultados demonstraram que a vimentina e a calponina foram percentualmente mais sensíveis que o HHF-35; entretanto, a calponina e o HHF-35 apresentaram padrão de distribuição focal diferentemente da distribuição difusa da vimentina. As células predominantemente marcadas, por todos os anticorpos utilizados, foram as não luminais presentes nas estruturas ductiformes e tubulares, vistas no adenoma pleomórfico e no carcinoma adenóide cístico, bem como as células dos cordões e ninhos dos adenocarcinomas polimorfo de baixo grau e periferia de lençóis e ninhos dos mioepiteliomas. Em conclusão, sugere-se que se faça associação da calponina com vimentina para identificação de células mioepiteliais em neoplasias de glândula salivar.