RESUMO
Thyroid cancer is maintaining at a high incidence level and its carcinogenesis is mainly affected by a complex gene interaction. By analysis of the next-generation resequencing of paired papillary thyroid cancer (PTC) and adjacent thyroid tissues, we found that Growth Associated Protein 43 (GAP43), a phosphoprotein activated by protein kinase C, might be novel markers associated with PTC. However, its function in thyroid carcinoma has been poorly understood. We discovered that GAP43 was significantly overexpressed in thyroid carcinoma and these results were consistent with that in The Cancer Genome Atlas (TCGA) cohort. In addition, some clinicopathological features of GAP43 in TCGA database showed that up-regulated GAP43 is significantly connected to lymph node metastasis (P < 0.001) and tumour size (P = 0.038). In vitro experiments, loss of function experiments was performed to investigate GAP43 in PTC cell lines (TPC-1 and BCPAP). The results proved that GAP43 knockdown in PTC cell significantly decreased the function of cell proliferation, colony formation, migration, and invasion and induced cell apoptosis. Furthermore, we also indicated that GAP43 could modulate the expression of epithelial-mesenchymal transition-related proteins, which could influence invasion and migration. Put those results together, GAP43 is a gene which was associated with PTC and might be a potential therapeutic target.
Assuntos
Proliferação de Células/genética , Transição Epitelial-Mesenquimal/genética , Proteína GAP-43/metabolismo , Metástase Linfática , Câncer Papilífero da Tireoide/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Adulto , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Estudos de Coortes , Bases de Dados Genéticas , Feminino , Proteína GAP-43/genética , Regulação Neoplásica da Expressão Gênica/genética , Técnicas de Silenciamento de Genes , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Estadiamento de Neoplasias , RNA Interferente Pequeno , Fatores de Risco , Câncer Papilífero da Tireoide/genética , Câncer Papilífero da Tireoide/patologia , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/secundárioRESUMO
In the present work, transgenic cucumber seedlings over expressing CsRCA and wild-type cucumber seedlings '08-1'at three-leaf stage exposed to high temperature (40 â, PFD 600 µmol· m-2 · s-1) were used to study the regulatory mechanism of photosynthesis by CsRCA. The results showed that the mRNA abundance of rbcL and rbcS as well as the activities of ribulose bisphosphate carboxylic enzyme (Rubisco) and Rubisco activase (RCA) were significantly higher in CsRCA over-expressing cucumber seedlings than in wild type (WT). Following 2-h exposure to high temperature, a notable decrease was observed in photosynthetic rate (Pn), photochemical perfor-mance index based on the absorption of light energy (PIABS), activities of Rubisco and RCA as well as the relative expression of rbcL, rbcS and CsRCA in both wild-type cucumber seedlings and transgenic cucumber seedlings. It was found that high temperature stress led to higher Wk, a parameter of chlorophyll (Chl) a fluorescence OJIP curve. Furthermore, high temperature greatly reduced the efficiency of electron transfer along the electron transport chain beyond QA(ψ0) and the quantum yield for electron transport (φE0), indicating that PSII oxygen complexes (OEC) and electron transport chain downstream QA were inhibited by high temperature. However, the inhibition could be alleviated by over expressing CsRCA in cucumber seedlings. Taken together, our data suggested that over expressing CsRCA improves photosynthesis in cucumber seedlings under high temperature stress by enhancing activities of the Rubisco and RCA, and maintaining the number of active reaction centers.
Assuntos
Cucumis sativus/fisiologia , Temperatura Alta , Fotossíntese , Proteínas de Plantas/genética , Clorofila , Clorofila A , Cucumis sativus/genética , Transporte de Elétrons , Plantas Geneticamente Modificadas/fisiologia , Ribulose-Bifosfato Carboxilase , Plântula , Estresse FisiológicoRESUMO
Taking the cucumber cultivar 'Jinyou 3' as test material, this paper studied the variations of the mRNA expression and activities of ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco), fructose-1, 6-bisphosphatase (FBPase), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), fructose-1, 6-bisphosphate aldolase (FBA), and transketolase (TK) in cucumber seedling leaves under suboptimal temperature and low light intensity (ST+LL). In the treatment of ST+LL, the leaf area and the dry mass per plant decreased remarkably, compared with the control. On the early days of ST+LL treatment, the gene expression of Rubisco rbcL and rbcS, FBPase, GAPDH, FBA, and TK declined markedly, the activities of the enzymes except TK obviously weakened, and the photosynthetic rate (P(n)) decreased rapidly. 3 days later, the gene expression of Rubisco rbcL and rbcS and the initial activity of Rubisco showed a continuous decrease but the decrement was obviously lesser, the total activity of Rubisco and the activities and gene expression of FBPase, GAPDH, FBA, and TK had an increasing trend, and the P(n) ascended simultaneously. When the treating time exceeded 6 days, the gene expression and the activities of Rubisco and FBPase tended to be constant, while those of the other enzymes as well as the P(n) presented a decreasing trend. These results suggested that the decline of the gene expression and activities of the photosynthetic enzymes in cucumber seedlings under suboptimal temperature and low light intensity was the important reason which led to the decrease of P(n). The adaptation of photosynthetic apparatus in cucumber seedlings to suboptimal temperature and low light intensity was related to the activation mechanisms of photosynthetic enzymes.
