SOX12 Promotes Thyroid Cancer Cell Proliferation and Invasion by Regulating the Expression of POU2F1 and POU3F1.

PURPOSE: SOX12 is overexpressed in many cancers, and we aimed to explore the biological function and mechanism of SOX12 in thyroid cancer. MATERIALS AND METHODS: We first analyzed the expression of SOX12 in thyroid cancer using data in The Cancer Genome Atlas. Immunohistochemistry and qRT-PCR were performed to identify SOX12 expression in thyroid cancer tissue and cells. Thyroid cancer cells were transfected with small interfering RNA targeting SOX12, and cellular functional experiments, including CCK8, wound healing, and Transwell assays, were performed. Protein expression was examined by Western blot analysis. A xenograft model was developed to evaluate the effect of SOX12 on tumor growth in vivo. RESULTS: SOX12 expression was increased in thyroid cancer tissue and cells. SOX12 promoted cell proliferation, migration, and invasion and accelerated tumor growth in vivo. The expression of PCNA, Cyclin D1, E-cadherin, Snail, MMP-2, and MMP-9 was affected by SOX12 knockdown. Bioinformatic analysis showed that SOX12 could interact with the POU family. SOX12 knockdown inhibited the expression of POU2F1, POU2F2, POU3F1 and POU3F2, and SOX12 expression showed a positive correlation with POU2F1, POU3F1, and POU3F2 expression in clinical data. POU2F1 and POU3F1 were able to reverse the effect of SOX12 knockdown on thyroid cancer cells. CONCLUSION: SOX12 affects the progression of thyroid cancer by regulating epithelial-mesenchymal transition and interacting with POU2F1 and POU3F1, which may be novel targets for thyroid cancer molecular therapy.

Marker-assisted pyramiding of QTLs for heat tolerance and escape upgrades heat resilience in rice (Oryza sativa L.).

KEY MESSAGE: This study demonstrated that pyramiding of early morning flowering and heat tolerance QTLs (qEMF3 and qHTSF4.1) in rice is an efficient approach to maintain high spikelet fertility under high-temperature stress at flowering stage. High temperature at flowering stage of rice causes low spikelet fertility and low yield. To cope with high-temperature stress brought by climate change, two strategies were proposed to develop heat-resilient rice varieties. One is to escape the high temperature by flowering early in the morning, another is to enhance tolerance to high-temperature stress per se. Two promising QTLs for early morning flowering (qEMF3) and heat tolerance (qHTSF4.1) were introgressed into IR64 background, and Near isogenic lines (NILs) IR64 + qEMF3 (IR64EMF3) and IR64 + qHTSF4.1 (IR64HT4) were developed in previous studies. In this study, a QTL pyramiding line IR64 + qHTSF4.1 + qEMF3 (IR64HT4EMF3) was developed by marker-assisted selection of the progenies of previous NILs. The NILs were subjected to different high-temperature regimes in the indoor growth chambers and different locations in the field. In the indoor growth chambers, when high temperature starts early (before 11:00 am), IR64HT4 and IR64HT4EMF3 had higher spikelet fertility than IR64EMF3; when high temperature comes later (after 11:00 am), IR64EMF3 and IR64HT4EMF3 had higher spikelet fertility than IR64HT4. The flowering pattern of the IR64HT4EMF3 was earlier than IR64HT4, but similar to IR64EMF3 in the glasshouse, field and indoor growth chambers. IR64HT4EMF3 showed higher spikelet fertility than IR64EMF3 and IR64HT4 in the field in the Philippines. Thus, combination of early morning flowering and heat tolerance QTLs is an elegant breeding strategy to cope with future extreme climate.

lncRNA FLVCR1-AS1 regulates cell proliferation, migration and invasion by sponging miR-485-5p in human cholangiocarcinoma.

Long non-coding RNA (lncRNA) FLVCR1 antisense RNA 1 (FLVCR1-AS1) serves a crucial role in many types of cancer; however, to the best of our knowledge, the biological effect of FLVCR1-AS1 in cholangiocarcinoma (CCA) remains unclear. The present study aimed to elucidate the involvement of FLVCR1-AS1 in the regulation of human CCA cell growth, migration and invasion, as well as the mechanisms underlying its effect. The expression levels of FLVCR1-AS1 in CCA tumor tissues, adjacent normal tissues, CCA cell lines and a cholangiocyte cell line were determined by reverse transcription-quantitative polymerase chain reaction. A significantly higher expression level of FLVCR1-AS1 was identified in CCA tumor tissues and the CCA cell lines HuCCT1 and CCLP1 compared with the normal controls. Short hairpin RNA targeting FLVCR1-AS1 (shFLVCR1-AS1) and a control plasmid (shNC) were transfected into CCA cell lines. Cell proliferation, colony formation, migration and invasion of CCA cells transfected with shFLVCR1-AS1 were significantly suppressed compared with the shNC groups. The expression levels of migration and invasion-associated proteins, including Twist, matrix metalloproteinase (MMP)-2 and MMP-9, were also significantly suppressed by shFLVCR1-AS1-treatment. Furthermore, FLVCR1-AS1 knockdown inhibited tumor growth in a xenograft model. Mechanistically, FLVCR1-AS1 was demonstrated to sponge microRNA-485-5p (miR-485-5p) in human CCA. The expression of miR-458-5p was significantly decreased in CCA tissue compared with normal tissue, and Pearson's correlation analysis revealed that FLVCR1-AS1 expression was negatively correlated with miR-485-5p expression in CCA tissues. These results suggested that lncRNA FLVCR1-AS1 may be used as a novel therapeutic target and a potential diagnostic marker for CCA.

KLF4 protects brain microvascular endothelial cells from ischemic stroke induced apoptosis by transcriptionally activating MALAT1.

In this study, we investigated the expression profile and functional role of Kruppel-like family of transcription factor 4 (KLF4) in mouse cerebral microvascular endothelial cells (MECs) after focal cerebral ischemia and in cultured b.End3 cells after oxygen-glucose deprivation (OGD). Its downstream regulation was also studied. Our data showed that KLF4 was significantly upregulated in the cerebral MECs after transient ischemic insult and in cultured b.End3 cells after 24 h OGD exposure. In in vitro b.End3 cell model, KLF4 shRNA significantly increased OGD-induced activation of caspase-3 and also increased OGD-induced b.End3 cell death. KLF4 shRNA substantially enhanced OGD induced Bim and Bax expression at mRNA and protein levels and also aggravated OGD induced upregulation of E-selectin, MCP-1 and IL-6. Bioinformatic analysis suggested that MALAT1 promoter has a possible KLF4 binding site. In this study, we validated this possible binding site and also demonstrated that enforced KLF4 expression increased MALAT1 transcription. Functionally, knockdown of MALAT1 phenocopied the effect of KLF4 shRNA on enhancing OGD induced cell apoptosis and OGD induced upregulation of pro-apoptotic factors and pro-inflammatory cytokines. Based on these findings, we infer that MALAT1 is a transcriptional target of KLF4 in its protective role in cerebral MECs after ischemic insult.

[Effects of different ecological conditions on grain quality and RVA profile of japonica rice from Yunnan of China and Korea].

Taking sixteen japonica rice varieties from Yunnan of China and Korea growing at three locations with different altitudes in Yunnan Plateau as test materials, this paper studied the effects of different ecological conditions on their grain quality, including brown rice length (BRL), brown rice width (BRW), length-width ratio (L/W), chalky rate (CR), whiteness (WH), amylose content (AC), protein content (PC), and alkali digestion value (ADV), and their starch RVA profile, including peak viscosity (PKV), hot viscosity (HTV), final viscosity (FLV), breakdown viscosity (BDV), setback viscosity (SBV), consistence viscosity( CTV), peak time (PeT), and pasting temperature (PaT). Of all the test parameters, SBV had the largest coefficient of variation. The coefficients of variation of BRL, CR, AC, PC, ADV, FLV, SBV, and PeT of Yunnan varieties were higher than those of Korean varieties, while the BRW, L/W, WH, PKV, HTV, BDV, CTV, and PaT of Korean varieties had higher coefficients of variation than those of Yunnan varieties. With increasing altitude, the BRL, BRW, L/W, WH, AC, ADV, FLV, and CTV of Yunnan varieties, and the BRL, BRW, WH, PC, PKV, HTV and BDV of Korean varieties decreased significantly, while the CR, PC, HTV, and PeT of Yunnan varieties, and the L/W, AC, ADV, SBV and CTV of Korean varieties increased significantly. The CR of Korean varieties had no obvious change. The PKV, BDV, and PaT of Yunnan varieties and the PaT of Korean varieties increased after an initial decrease, whereas the SBV of Yunnan varieties and the FLV and PeT of Korean varieties were in reverse.