MicroRNA-524-5p increases cisplatin-sensitivity of gastric cancer cells by targeting SOX9 gene / 肿瘤

Objective: To investigate the mechanism of microRNA (miR)-524-5p mediating the effect of cisplatin in the treatment of gastric cancer. Methods: The level of miR-524-5p in gastric cancer tissues, the matched non-tumor tissues, cisplatin-resistant and parental gastric cancer cell lines was measured by real-time fluorescent quantitative PCR. The viability of cisplatin-resistant gastric cancer SGC-7901 and MKN-45 cells transfected with miR-524-5p mimics or the control RNA was detected by CCK-8 method. The expression of SRY-related high mobility group-box gene 9 (SOX9), a targeting protein of miR-524-5p, was identified by luciferase reporter assay and Western blotting. The “rescue” assay was used to explore the effect of SOX9 overexpression on the cisplatin-resistance of gastric cancer cells induced by miR-524-5p. Results: Compared with the normal gastric epithelial GES1 cells, the level of miR-524-5p in gastric cancer SGC-7901 and MKN-45 cells was significantly down-regulated (both P < 0.05); while the level of miR-524-5p in gastric cancer tissues was significantly lower than that in the adjacent non-tumor tissues (P < 0.05). After the treatment of cisplatin, the expression level of miR-524-5p in cisplatin-resistant gastric cancer SGC-7901 and MKN-45 cells was decreased as compared with the sensitive parental cells (both P< 0.05). After the transfection of miR-524-5p mimics, the sensitivity of cisplatin-resistant gastric cancer SGC-7901 and MKN-45 cells was significantly enhanced (both P< 0.05). Subsequently, SOX9 was identified as a functional target gene of miR-524-5p. Overexpression of SOX9 could counteract the chemosensitizing effect of miR-524-5p, in other words, the cisplatin-sensitivity of SGC-7901 and MKN-45 cells transfected with SOX9 overexpression plasmids and miR-524-5p mimics was significantly lower than that in single miR-524-5p mimics transfection group (both P< 0.05). Conclusion: The expression of miR-524-5p influences the sensitivity of human gastric cancer cells to cisplatin, suggesting that miR-524-5p has the potential to be a novel target for the treatment of chemoresistant gastric cancer patients.

PLAG1, SOX10, and Myb Expression in Benign and Malignant Salivary Gland Neoplasms

BACKGROUND: Recent findings in molecular pathology suggest that genetic translocation and/or overexpression of oncoproteins is important in salivary gland tumorigenesis and diagnosis. We investigated PLAG1, SOX10, and Myb protein expression in various salivary gland neoplasm tissues. METHODS: A total of 113 cases of surgically resected salivary gland neoplasms at the National Cancer Center from January 2007 to March 2017 were identified. Immunohistochemical staining of PLAG1, SOX10, and Myb in tissue samples was performed using tissue microarrays. RESULTS: Among the 113 cases, 82 (72.6%) were benign and 31 (27.4%) were malignant. PLAG1 showed nuclear staining and normal parotid gland was not stained. Among 48 cases of pleomorphic adenoma, 29 (60.4%) were positive for PLAG1. All other benign and malignant salivary gland neoplasms were PLAG1-negative. SOX10 showed nuclear staining. In normal salivary gland tissues SOX10 was expressed in cells of acinus and intercalated ducts. In benign tumors, SOX10 expression was observed in all pleomorphic adenoma (48/48), and basal cell adenoma (3/3), but not in other benign tumors. SOX10 positivity was observed in nine of 31 (29.0%) malignant tumors. Myb showed nuclear staining but was not detected in normal parotid glands. Four of 31 (12.9%) malignant tumors showed Myb positivity: three adenoid cystic carcinomas (AdCC) and one myoepithelial carcinoma with focal AdCC-like histology. CONCLUSIONS: PLAG1 expression is specific to pleomorphic adenoma. SOX10 expression is helpful to rule out excretory duct origin tumor, but its diagnostic value is relatively low. Myb is useful for diagnosing AdCC when histology is unclear in the surgical specimen.

Expression and mechanism of microRNA-613 in breast cancer / 中国肿瘤临床

ABSTRACT Objective: To investigate the expression and mechanism of microRNA (miRNA)-613 in breast cancer. Methods: A total of 91 specimens of breast cancer tissue were collected from Nanchong Central Hospital between May 2017 and May 2018. Quantitative real-time PCR (qRT-PCR) was used to estimate miRNA-613 expression levels in breast cancer and adjacent tissues and breast cancer ( cells MDA-MB-231, MDA-MB-468, and MCF-7) and normal breast epithelial (HBL-100) cell lines. Based on these data, the relationship between miRNA-613 expression and clinicopathological characteristics and prognosis in breast cancer patients were analyzed using the cancer genome atlas (TCGA) database. A dual-luciferase reporter assay was used to detect the binding of miRNA-613 to the 3'UTR of SOX9. Effects on cell proliferation and cell invasion and migration upon transfection of MDA-MB-231 cells with miRNA-613 mimics were detected by the CCK-8 assay and Transwell invasion and migration assays, respectively. In addition, Western blot was used to estimate the expression levels of SOX9, β-catenin, E-cadherin, and Vimentin in the transfected cells. Results: The expression of miRNA-613 in breast cancer tissues was significantly lower than that in adjacent tissues (P<0.05) and was found to be closely related to TNM stage and lymph node metastasis (P<0.05). TCGA survival data showed that miRNA-613 expression was not related to the overall survival rate of breast cancer patients (P>0.05 ). The expression of miRNA-613 in breast cancer cell lines was significantly lower than that in the normal breast epithelial cell line (P<0.05). Similarly, the expression of miRNA-613 in highly invasive metastatic breast cancer cell lines (MDA-MB-231 and MDA-MB-468) was significantly lower than that in the metastatic breast cancer cell line MCF-7 with lower invasion ability (P<0.05). The dual-luciferase reporter assay showed that miRNA-613 could specifically bind to the 3'UTR of SOX9. Upregulation of miRNA-613 expression could inhibit the proliferation, invasion, and migration of MDA-MB-231 cells (P<0.05). This was associated with the downregulated expression of SOX9, β-catenin, and Vimentin (P<0.05) and upregulation of E-Cadherin expression (P<0.05). Conclusions: The expression of miRNA-613 was decreased in breast cancer tissues and cell lines. MiRNA-613 may inhibit the proliferation, invasion, and metastasis of breast cancer cells and epithelial-mesenchymal transition by regulating the SOX9 and Wnt/β-catenin signaling pathways.

Clinicopathologic features of mammary analogue secretory carcinoma of salivary glands / 中华病理学杂志

Objective@#To investigate the clinicopathological features of mammary analogue secretory carcinoma (MASC) of salivary glands, and its diagnosis, differential diagnosis, immunohistochemistry and molecular pathology.@*Methods@#Seventeen cases of MASC were enrolled, with 9 cases of salivary acinar cell carcinoma and 18 cases of adenoid cystic carcinoma as control groups from Nanjing General Hospital from 1997 to 2014 were included in this retrospective study, combined with immunohistochemistry and molecular detection of ETV6-NTRK3 gene fusion. All cases were histologically reviewed with immunohistochemical staining (EnVision) for S-100 protein, SOX10, GATA3, CD117 expression in each group. Fluorescence in situ hybridization (FISH) was used to detect the ETV6-NTRK3 gene fusion.@*Results@#The age of MASC patients ranged from 27 to 74 years with mean age of 47 and ratio of male and female was 4∶3. All cases showed infiltrative growth and diverse cytology and histology, including lobular (8 cases), cystic papillary (3 cases), cribriform mixed with papillary and glandular structures (6 cases) at various proportions. Some tumors of MASC also exhibited solid growth areas with occasional microcystic honeycombed pattern composed of small cysts merged into larger cysts resembling thyroid follicles. S-100 protein and SOX10 were strongly positive in all MASC cases (17/17). In addition, there was insignificant positivity for GATA3 (3/17) and CD117 (4/17). ETV6 gene fusion detection was informative in 12 MASC cases by FISH with 10 positive cases and 2 negative cases.@*Conclusions@#Combined immunohistochemical positivity of S-100 protein, CD117 and SOX10 are useful in the diagnosis and differential diagnosis of MASC. FISH detection of ETV6-NTRK3 fusion offers an additional molecular diagnostic marker for the diagnosis.