ABSTRACT
OBJECTIVE: Selecting stably expressed reference genes is crucial for evaluating real-time quantitative polymerase chain reaction (RT-qPCR) data via the relative quantification method. In the present-day study, our aim was to select optimal reference genes (RGs) for the investigation of target gene (TG) expression profiling in cancerous human laryngeal and hypopharyngeal tissues. PATIENTS AND METHODS: 12 cancerous laryngeal tissues and 10 cancerous hypopharyngeal tissues were investigated. The expression characteristics of 11 reference genes (18S rRNA, GAPDH, B2M, ACTB, TBP, ALAS1, RPL29, HMBS, HPRT1, GUSB, and PUM1), which were commonly used in RT-qPCR for the analysis of gene expression, were investigated using the geNorm, NormFinder, and BestKeeper algorithm programs. RESULTS: HMBS, ALAS1, and B2M were suggested as optimal RGs for studying human laryngeal and hypopharyngeal cancerous tissues together, laryngeal cancerous tissue by itself, and hypopharyngeal cancerous tissue by itself, respectively. If 2 or more reference genes are needed to achieve better standardization, 3 reference genes can optimally be used in combination to improve the accuracy of relative quantitation normalization. The recommended combinations for studying human laryngeal and hypopharyngeal cancerous tissues together, laryngeal cancerous tissue by itself, and hypopharyngeal cancerous tissue by itself were HMBS + HPRT1 + GUSB, ALAS1 + GUSB + HMBS, and B2M + HPRT1 + TBP, respectively. CONCLUSIONS: The recommended reference genes could be used to improve the accuracy of gene expression studies on the molecular mechanisms of cancerous human laryngeal and hypopharyngeal tissues. The selected combination of reference genes can effectively improve the accuracy of the relative quantitative diagnosis of gene expression levels, such as messenger RNA, circular RNA, and long-noncoding RNA.
Subject(s)
Hypopharyngeal Neoplasms/genetics , Laryngeal Neoplasms/genetics , Real-Time Polymerase Chain Reaction/methods , Aged , Algorithms , China/epidemiology , Evaluation Studies as Topic , Female , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic , Humans , Hypopharyngeal Neoplasms/pathology , Laryngeal Neoplasms/pathology , Male , Middle Aged , Neoplasm Staging/methods , RNA, Messenger/geneticsABSTRACT
A case of an unusually large sialolith arising in the parenchyma of submandibular gland accompany with infection is presented, computered tomography identified a sialolith in the submandibular gland. Surgery on the sialolith was subsequently completed under general anesthesia extraorally. A brownish stone was present in the parenchyma of the submandibular gland, measuring 33 mm×25 mm.
Subject(s)
Salivary Gland Calculi/complications , Soft Tissue Infections/complications , Submandibular Gland Diseases/complications , Humans , Submandibular GlandABSTRACT
OBJECTIVE: Increasing evidence indicates that MicroRNAs, a class of small RNA molecules, play crucial roles in tumorigenesis, through affecting cell proliferation, apoptosis and metastasis. The present study aimed to investigate the effect of miR-205 on gastric cancer cell proliferation. MATERIALS AND METHODS: The expression of miR-205 was examined in the gastric cancer tissues and cell lines. BrdU incorporation assay was used to measure the cell proliferation. Western blot was performed to determine the protein expression. RESULTS: miR-205 is significantly down-regulated in gastric cancer tissues, compared with adjacent normal tissues. Besides, miR-205 expression is associated with clinical and pathological characteristics of patients. In vitro studies further found that inhibition of miR-205 significantly promoted gastric cancer cell proliferation via cell-cycle progression. Further analyses indicated that miR-205 was able to repress oncoprotein Yin Yang 1 expression, through targeting its 3'-untranlated region. CONCLUSIONS: Our data suggest that down-regulation of miR-205 may represent an important mechanism for the development of gastric cancer.
