目的探讨长链非编码RNA ABHD11-AS1对胃癌细胞糖酵解的影响及其作用的分子机制。方法分别将空载质粒pcDNA-Vector和过表达质粒pcDNA-ABHD11-AS1转染入ABHD11-AS1低表达的MKN45和MGC803胃癌细胞中,构建过表达组(pcDNA-ABHD11-AS1)和空载...目的探讨长链非编码RNA ABHD11-AS1对胃癌细胞糖酵解的影响及其作用的分子机制。方法分别将空载质粒pcDNA-Vector和过表达质粒pcDNA-ABHD11-AS1转染入ABHD11-AS1低表达的MKN45和MGC803胃癌细胞中,构建过表达组(pcDNA-ABHD11-AS1)和空载质粒对照组胃癌细胞株。运用CCK-8法检测细胞的增殖活性并绘制生长曲线,克隆形成实验检测细胞克隆形成能力,Transwell检测细胞迁移、侵袭能力,葡萄糖摄取实验及乳酸生成实验检测细胞糖酵解水平的变化;LncMAP数据库分析查找ABHD11-AS1可能调控的转录因子,查阅文献进行分析挑选候选转录因子,Western blot明确ABHD11-AS1是否影响候选转录因子的表达量。结果与对照组相比,转染pcDNA-ABHD11-AS1后,MGC803和MKN45胃癌细胞中ABHD11-AS1基因表达量明显上升(P<0.01),CCK8和成克隆实验表明细胞增殖加快(P<0.05),克隆形成能力增强,Tanswell实验结果证实细胞迁移(11±2 vs 27±3;17±4 vs 28±3,P<0.01)、侵袭(15±3 vs 26±2;10±1 vs 35±2,P<0.01)作用增强;上调ABHD11-AS1后,胃癌细胞葡萄糖摄取及乳酸生成增多(P<0.05)。分析数据库结果显示ABHD11-AS1可能调控经典糖酵解相关基因c-Myc,Western blot结果证实上调ABHD11-AS1后,c-Myc表达量随之升高。结论ABHD11-AS1通过上调c-Myc促进胃癌细胞内的糖酵解,并加速胃癌进展。展开更多
BACKGROUND Gastric cancer (GC) is the fourth most frequent malignancy all over the world. The diagnosis of GC is challenging and the prognosis of GC is very unfavorable. Accumulating evidence reveals that serum long n...BACKGROUND Gastric cancer (GC) is the fourth most frequent malignancy all over the world. The diagnosis of GC is challenging and the prognosis of GC is very unfavorable. Accumulating evidence reveals that serum long noncoding RNAs (lncRNAs) can function as biomarkers in various types of cancers, including GC. AIM To explore the level and molecular mechanism of the lncRNA HOXA11-AS in GC and the diagnostic and prognostic significance of serum HOXA11-AS in GC. METHODS HOXA11-AS levels in GC tissue, cell lines, and serum samples were measured. The correlation between HOXA11-AS expression and clinicopathological characteristics was analyzed. The role of HOXA11-AS in the diagnosis and prognosis of GC was evaluated. Cell function assays were performed for exploration of the roles of HOXA11-AS in GC cells. Moreover, Western blot was performed to explore the target regulated by HOXA11-AS in GC cells. RESULTS Up-regulation of HOXA11-AS was found in GC tissues, cell lines, and serum samples. In GC patients, decreased serum HOXA11-AS levels were negatively related with tumor size, TNM stage, and lymph node metastasis. The area under the receiver operating characteristic curve of serum HOXA11-AS in the diagnosis of GC was 0.924 (95%CI: 0.881-0.967;sensitivity, 0.787;specificity 0.978). Results of the Kaplan-Meier survival curves suggested the GC patients with a lower HOXA11-AS level having a better overall survival rate. HOXA11-AS promoted GC cell proliferation and invasion. SRSF1 may be the target regulated by HOXA11-AS in GC cells. CONCLUSION HOXA11-AS promotes GC cell proliferation and invasion via SRSF1 and may function as a promising marker in GC.展开更多
Long noncoding RNAs(lnc RNAs)participate in many pathophysiological processes after traumatic brain injury by mediating neuroinflammation and apoptosis.Homeobox A11 antisense RNA(HOXA11-AS)is a member of the lnc RNA f...Long noncoding RNAs(lnc RNAs)participate in many pathophysiological processes after traumatic brain injury by mediating neuroinflammation and apoptosis.Homeobox A11 antisense RNA(HOXA11-AS)is a member of the lnc RNA family that has been reported to participate in many inflammatory reactions;however,its role in traumatic brain injury remains unclear.In this study,we established rat models of traumatic brain injury using a weight-drop hitting device and injected LV-HOXA11-AS into the right lateral ventricle 2 weeks before modeling.The results revealed that overexpression of HOXA11-AS aggravated neurological deficits in traumatic brain injury rats,increased brain edema and apoptosis,promoted the secretion of proinflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factorα,and promoted the activation of astrocytes and microglia.Microglia were treated with 100 ng/m L lipopolysaccharide for 24 hours to establish in vitro cell models,and then transfected with pc DNA-HOXA11-AS,mi R-124-3 p mimic,or sh-MDK.The results revealed that HOXA11-AS inhibited mi R-124-3 p expression and boosted MDK expression and TLR4-nuclear factor-κB pathway activation.Furthermore,lipopolysaccharide enhanced potent microglia-induced inflammatory responses in astrocytes.Forced overexpression of mi R-124-3 p or downregulating MDK repressed microglial activation and the inflammatory response of astrocytes.However,the mi R-124-3 p-mediated anti-inflammatory effects were reversed by HOXA11-AS.These findings suggest that HOXA11-AS can aggravate neuroinflammation after traumatic brain injury by modulating the mi R-124-3 p-MDK axis.This study was approved by the Animal Protection and Use Committee of Southwest Medical University(approval No.SMU-2019-042)on February 4,2019.展开更多
文摘目的探讨长链非编码RNA ABHD11-AS1对胃癌细胞糖酵解的影响及其作用的分子机制。方法分别将空载质粒pcDNA-Vector和过表达质粒pcDNA-ABHD11-AS1转染入ABHD11-AS1低表达的MKN45和MGC803胃癌细胞中,构建过表达组(pcDNA-ABHD11-AS1)和空载质粒对照组胃癌细胞株。运用CCK-8法检测细胞的增殖活性并绘制生长曲线,克隆形成实验检测细胞克隆形成能力,Transwell检测细胞迁移、侵袭能力,葡萄糖摄取实验及乳酸生成实验检测细胞糖酵解水平的变化;LncMAP数据库分析查找ABHD11-AS1可能调控的转录因子,查阅文献进行分析挑选候选转录因子,Western blot明确ABHD11-AS1是否影响候选转录因子的表达量。结果与对照组相比,转染pcDNA-ABHD11-AS1后,MGC803和MKN45胃癌细胞中ABHD11-AS1基因表达量明显上升(P<0.01),CCK8和成克隆实验表明细胞增殖加快(P<0.05),克隆形成能力增强,Tanswell实验结果证实细胞迁移(11±2 vs 27±3;17±4 vs 28±3,P<0.01)、侵袭(15±3 vs 26±2;10±1 vs 35±2,P<0.01)作用增强;上调ABHD11-AS1后,胃癌细胞葡萄糖摄取及乳酸生成增多(P<0.05)。分析数据库结果显示ABHD11-AS1可能调控经典糖酵解相关基因c-Myc,Western blot结果证实上调ABHD11-AS1后,c-Myc表达量随之升高。结论ABHD11-AS1通过上调c-Myc促进胃癌细胞内的糖酵解,并加速胃癌进展。
文摘BACKGROUND Gastric cancer (GC) is the fourth most frequent malignancy all over the world. The diagnosis of GC is challenging and the prognosis of GC is very unfavorable. Accumulating evidence reveals that serum long noncoding RNAs (lncRNAs) can function as biomarkers in various types of cancers, including GC. AIM To explore the level and molecular mechanism of the lncRNA HOXA11-AS in GC and the diagnostic and prognostic significance of serum HOXA11-AS in GC. METHODS HOXA11-AS levels in GC tissue, cell lines, and serum samples were measured. The correlation between HOXA11-AS expression and clinicopathological characteristics was analyzed. The role of HOXA11-AS in the diagnosis and prognosis of GC was evaluated. Cell function assays were performed for exploration of the roles of HOXA11-AS in GC cells. Moreover, Western blot was performed to explore the target regulated by HOXA11-AS in GC cells. RESULTS Up-regulation of HOXA11-AS was found in GC tissues, cell lines, and serum samples. In GC patients, decreased serum HOXA11-AS levels were negatively related with tumor size, TNM stage, and lymph node metastasis. The area under the receiver operating characteristic curve of serum HOXA11-AS in the diagnosis of GC was 0.924 (95%CI: 0.881-0.967;sensitivity, 0.787;specificity 0.978). Results of the Kaplan-Meier survival curves suggested the GC patients with a lower HOXA11-AS level having a better overall survival rate. HOXA11-AS promoted GC cell proliferation and invasion. SRSF1 may be the target regulated by HOXA11-AS in GC cells. CONCLUSION HOXA11-AS promotes GC cell proliferation and invasion via SRSF1 and may function as a promising marker in GC.
基金supported by the Science and Technology Project of Sichuan Province of China,No.2020YJ0188the Science and Technology Foundation of Luzhou of China,No.2017LZXNYD-J10(both to XLL)。
文摘Long noncoding RNAs(lnc RNAs)participate in many pathophysiological processes after traumatic brain injury by mediating neuroinflammation and apoptosis.Homeobox A11 antisense RNA(HOXA11-AS)is a member of the lnc RNA family that has been reported to participate in many inflammatory reactions;however,its role in traumatic brain injury remains unclear.In this study,we established rat models of traumatic brain injury using a weight-drop hitting device and injected LV-HOXA11-AS into the right lateral ventricle 2 weeks before modeling.The results revealed that overexpression of HOXA11-AS aggravated neurological deficits in traumatic brain injury rats,increased brain edema and apoptosis,promoted the secretion of proinflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factorα,and promoted the activation of astrocytes and microglia.Microglia were treated with 100 ng/m L lipopolysaccharide for 24 hours to establish in vitro cell models,and then transfected with pc DNA-HOXA11-AS,mi R-124-3 p mimic,or sh-MDK.The results revealed that HOXA11-AS inhibited mi R-124-3 p expression and boosted MDK expression and TLR4-nuclear factor-κB pathway activation.Furthermore,lipopolysaccharide enhanced potent microglia-induced inflammatory responses in astrocytes.Forced overexpression of mi R-124-3 p or downregulating MDK repressed microglial activation and the inflammatory response of astrocytes.However,the mi R-124-3 p-mediated anti-inflammatory effects were reversed by HOXA11-AS.These findings suggest that HOXA11-AS can aggravate neuroinflammation after traumatic brain injury by modulating the mi R-124-3 p-MDK axis.This study was approved by the Animal Protection and Use Committee of Southwest Medical University(approval No.SMU-2019-042)on February 4,2019.
