TM2D1 contributes the epithelial-mesenchymal transition of hepatocellular carcinoma via modulating AKT/β-catenin axis

Various epidemiology studies showed the correlation between Alzheimer’s disease (AD) and low incidence of cancer. However, the etiology underlying etiology of AD-related carcinogenesis remains largely elusive. Our study focused on characterizing the role of TM2D1 (TM2 domain containing 1) in hepatocellular carcinoma. TM2D1 is also known as β-amyloid peptide binding protein and is critical to the pathogenesis of AD. We found that TM2D1 is increasingly expressed in HCC tumors relative to the peritumoral tissues of the matched tumors and high TM2D1 expression predicts unfavorable clinical outcomes. TM2D1 overexpression induced HCC cell proliferation, migration and invasion, which was related to the epithelial-mesenchymal transition (EMT) observed in these cells. Conversely, TM2D1 depletion led to opposite phenotype in HCC. Mechanistically, we found that TM2D1 promoted Akt and β-catenin hyper-activation, which corresponded with molecular marker change in EMT signaling pathway. Taken together, our results indicated that TM2D1 played an important role in the EMT process in HCC cells by activating AKT and β-catenin signaling and may become a promising therapeutic target in HCC.     

SETDB1 in cancer: overexpression and its therapeutic implications

SET Domain Bifurcated Histone Lysine Methyltransferase 1 (SETDB1, ESET, KMT1E) is a H3K9 methyltransferase involved in gene silencing. In recent years, SETDB1 has been implicated as an oncogene in various cancers, highlighting a critical need to better understand the mechanisms underlying SETDB1 amplification, overexpression, and activation. In the following review, we first examine the history of SETDB1, starting from its discovery in 1999 and ending with recent findings. We follow with an outline of the structure and subcellular location of SETDB1, as well as potential mechanisms for regulation of its nuclear transport. Subsequently, we introduce SETDB1’s various functions, including its roles in promyelocytic leukemia nuclear body (PML-NB) formation, the methylation and activation of Akt, the silencing of the androgen receptor (AR) gene, retroelement silencing, the inhibition of tumor suppressor p53, and its role in promoting intestinal differentiation and survival. The Cancer Cell Line Encyclopedia (CCLE) screened SETDB1 dependency in 796 cancer cell lines, identifying SETDB1 as a common essential gene in 531 of them, demonstrating that SETDB1 expression is critical for the survival of the majority of cancers. Therefore, we provide a detailed review of the oncogenic effects of SETDB1 overexpression in breast cancer, non-small cell lung cancer, prostate cancer, colorectal cancer, acute myeloid leukemia, glioma, melanoma, pancreatic ductal adenocarcinoma, liver cancer, nasopharyngeal carcinoma, gastric carcinoma, and endometrial cancer. Accordingly, we review several methods that have been used to target SETDB1, such as using Mithramycin A, Mithralog EC-8042, 3’-deazaneplanocin A (DZNep), and paclitaxel. Finally, we conclude by highlighting remaining gaps in knowledge and challenges surrounding SETDB1. Ultimately, our review captures the wide scope of findings on SETDB1’s history, function, its implications in cancer, and provides suggestions for future research in the field.     

AKIP1 promotes glioblastoma viability,mobility and chemoradiation resistance via regulating CXCL1 and CXCL8 mediated NF-κB and AKT pathways

This study aimed to investigate the interaction of A-kinase-interacting protein 1 (AKIP1) with C-X-C motif chemokine ligand (CXCL)1, CXCL2, CXCL8, and their effects on regulating glioblastoma multiforme (GBM) malignant behaviors. AKIP1 expression was modified by pcDNA and pGPH1 vectors in U-87 MG and U-251 MG cells. Subsequently, multiple compensative experiments were conducted via adding CXCL1, CXCL2 and CXCL8 in the pGPH1-AKIP1 (AKIP1 knockdown) transfected U-87 MG and U-251 MG cells, respectively. Furthermore, AKIP1, CXCL1/2/8 expressions in 10 GBM and 10 low-grade glioma (LGG) tumor samples were detected. AKIP1 was elevated in various GBM cell lines compared to normal human astrocytes. AKIP1 overexpression promoted U-87 MG and U-251 MG cell proliferation and invasion while inhibited apoptosis; and it enhanced chemoresistance to temozolomide (but not cisplatin) and radiation resistance; then AKIP1 knockdown showed the opposite effects. Meanwhile, AKIP1 positively regulated CXCL1/2/8, NF-κB pathway, AKT pathway and PD-L1 expression. Further multiple compensative experiments uncovered that CXCL1 and CXCL8 promoted proliferation, invasion, chemoradiation resistance, NF-κB pathway, AKT pathway and PD-L1 expression in U-87 MG and U-251 MG cells, also in pGPH1-AKIP1 (AKIP1 knockdown) transfected U-87 MG and U-251 MG cells; although CXCL2 exhibited similar treads, but its effect was much weaker. Besides, NF-κB pathway inhibitor and AKT pathway inhibitor attenuated the effect of CXCL1&CXCL8 on promoting GBM cell malignant behaviors. Clinically AKIP1 and CXCL1/8 were elevated in GBM compared to LGG tumor samples, and they were inter-correlated. AKIP1 promotes GBM viability, mobility and chemoradiation resistance via regulating CXCL1 and CXCL8 mediated NF-κB and AKT pathways.     

乐伐替尼靶向调节VEGFR/PI3K/AKT治疗晚期甲状腺癌的作用机制

目的探讨乐伐替尼靶向调节VEGFR/PI3K/AKT治疗晚期甲状腺癌的作用机制。方法选取100例晚期分化型甲状腺癌患者,采用口服乐伐替尼,剂量为24 mg/d,直到出现无法耐受的副作用和疾病恶化。评价药物使用疗效;观察患者甲状腺癌血清标志物Tg水平变化;采用免疫组织化学法检测甲状腺癌患者癌组织中VEGFR2蛋白的表达;采用Western Blot法和Real-Time PCR法检测甲状腺癌癌组织中PI3K、AKT的磷酸化蛋白和基因水平;采用Caspase 3试剂盒测定凋亡蛋白Caspase 3表达。结果乐伐替尼治疗晚期分化型甲状腺癌的有效率为60.00%,患者血清TgAb浓度较治疗前明显下降(P<0.05)。与癌旁组织相比,甲状腺癌组织内Caspase 3活性、VEGFR2蛋白表达含量、p-PI3K和p-AKT蛋白含量及PI3K和AKT mRNA表达明显上升(P<0.05)。晚期分化型甲状腺癌患者应用乐伐替尼治疗后,Caspase 3活性、VEGFR2蛋白表达含量、p-PI3K和p-AKT蛋白含量及PI3K和AKT mRNA表达较治疗前显著降低,差异具有统计学意义(P<0.05)。结论乐伐替尼可以有效治疗晚期分化型甲状腺癌,并调控VEGFR靶向途径抑制PI3K/AKT信号通路,诱导甲状腺癌细胞凋亡。     

PI3K/AKT/mTOR信号通路抑制剂在卵巢癌治疗中的研究进展

卵巢癌是女性生殖系统最致命的恶性肿瘤。目前,针对卵巢癌的规范治疗方案是肿瘤细胞减灭术辅以紫杉醇/铂类联合化疗,然而大多数晚期卵巢癌患者最终因对化疗药物耐药而复发。PI3K/AKT/m TOR信号通路作为一条重要的原癌基因通路,在卵巢癌中激活并在卵巢癌的增殖、侵袭、细胞周期进程、血管形成及耐药中发挥着重要的作用,抑制该通路是卵巢癌的一个潜在治疗方法。对PI3K/AKT/m TOR信号通路抑制剂在卵巢癌治疗中的研究进展进行综述。     

SET8介导AKT信号通路在调控高磷诱导的血管平滑肌细胞钙化中的作用

目的 探讨赖氨酸甲基转移酶SET8介导AKT信号通路在调控高磷诱导的血管平滑肌细胞(vascular smooth muscle cells, VSMCs)钙化中的作用及机制。方法 (1)选取雄性SD大鼠进行体内实验并随机分为假手术组和慢性肾衰竭血管钙化组。取胸主动脉,采用von Kossa染色检测钙化情况;免疫组化法检测SET8和Caspase-3表达情况。(2)体外实验进行原代VSMCs培养,将细胞随机分为正常组和高磷组(10 mmol/L β-甘油磷酸)。采用邻甲酚酞络合酮比色法及茜素红染色检测细胞钙化情况;流式细胞法检测细胞凋亡情况;RT-PCR和Western印迹法检测SET8、AKT和Caspase-3的表达。(3)为进一步验证SET8在VSMCs凋亡中的作用,采用脂质体转染法,分3组:SET8-短发夹RNA(shRNA)组、空质粒组和正常对照组。采用邻甲酚酞络合酮比色法及茜素红染色检测细胞钙化情况;流式细胞法检测细胞凋亡情况;RT-PCR和Western印迹法检测SET8、AKT、Caspase-3的表达。结果 (1)体内实验中,慢性肾衰竭血管钙化组的血管钙盐沉积量明显高于假手术组(P<0.05);免疫组化结果提示,与假手术组相比,血管钙化组SET8表达明显降低,Caspase-3表达明显升高(均P<0.05);相关分析显示,血管组织SET8表达水平与钙含量、Caspase-3表达呈负相关(r=-0.948,P<0.01;r=-0.961,P<0.01)。(2)体外实验中,高磷组钙盐沉积明显高于正常组(P<0.05);流式细胞术检测结果显示,高磷组VSMCs的凋亡率明显高于正常组(P<0.05);RT-PCR和Western印迹显示,与正常组相比,高磷组SET8和AKT的mRNA和蛋白表达下降,Caspase-3的mRNA和蛋白表达升高(均P<0.05)。(3)干扰SET8基因表达后,VSMCs钙化及凋亡率明显增加,AKT的mRNA和蛋白表达下降,Caspase-3的mRNA和蛋白表达升高(均P<0.05)。结论 SET8可以抑制血管钙化,其机制之一可能为通过促进AKT活化,抑制Caspase-3的表达,从而抑制VSMCs凋亡,进而参与调控高磷诱导的血管平滑肌细胞钙化。     

基于CaSR表达及AKT磷酸化水平的花旗泽仁改善胰岛素抵抗作用机制

目的基于CaSR表达及AKT磷酸化水平考察花旗泽仁改善胰岛素抵抗的作用机制。方法采用高浓度胰岛素体外诱导培养法建立IR L02肝细胞模型,并通过葡萄糖消耗实验对细胞模型进行鉴定。采用免疫荧光法和qRT-PCR技术检测CaSR表达的细胞定位以及CaSR mRNA表达,同时采用Western blot技术检测CaSR蛋白、磷酸化AKT(Ser473和Thr308)蛋白表达。结果高浓度胰岛素诱导后的L02肝细胞模型葡萄糖消耗量和摄取率明显下降(P<0.01)。免疫荧光实验结果表明,模型对照组的CaSR分布较稀疏,相对应的,给予花旗泽仁后的CaSR分布较密集。通过qRT-PCR实验发现,模型对照组的CaSR mRNA表达显著下调(P<0.01);与模型对照组比较,花旗泽仁组、阳性对照组中CaSR mRNA表达明显上调(P<0.01)。同时Western blot检测结果表明,模型对照组的CaSR和磷酸化AKT(Thr308和Ser473)蛋白表达显著下降(P<0.01),与模型对照组比较,花旗泽仁组、阳性对照组中CaSR和磷酸化AKT(Thr308和Ser473)蛋白表达明显升高(P<0.01,P<0.05)。结论该实验证明了花旗泽仁通过影响PI3K通路改善胰岛素抵抗,可能与通过调节CaSR,激活PI3K/AKT信号通路AKT上的2个(Thr308,Ser473)磷酸化位点的蛋白表达有关。     

网络药理学探讨淫羊藿-巴戟天药对治疗勃起功能障碍的潜在机制

目的：运用网络药理学的分析方法探讨淫羊藿-巴戟天治疗勃起功能障碍的潜在机制。方法：从中药系统药理学数据库与分析平台（TCMSP）、生物信息学分析工具BATMAN-TCM数据库筛选出淫羊藿、巴戟天的有效活性成分及作用靶点,运用GeneCards、OMIM、NCBI数据库获取勃起功能障碍的疾病靶点,韦恩图筛选药对、疾病的共同的靶点。采用String数据库进行蛋白质-蛋白质相互作用（PPI）网络构建,Cystoscap 372进行拓扑分析,构建药对中药-活性成分-疾病-靶点交集网络,运用R363软件对共同靶点进行基因本体（GO）富集分析和京都基因和基因组百科全书（KEGG）富集分析,筛选出潜在信号通路并分析其作用机制。结果：1）巴戟天、淫羊藿筛选出203个靶点。2）勃起功能障碍疾病筛选出1 004个靶点。3）淫羊藿-巴戟天药对与勃起功能障碍的交集靶点共75个。4）通过PPI网络筛选出IL6、VEGFA、CASP3等30个关键靶点。5）GO分析的生物过程主要富集在对脂多糖的反应等过程;细胞组分主要富集在膜筏、膜微区等部位;分子功能主要富集在肾上腺素能受体活性等。6）KEGG分析共富集到PI3K-AKT信号通路等17条信号通路。结论：淫羊藿-巴戟天药对治疗勃起功能障碍,主要通过影响PI3K-AKT等信号通路发挥作用,同时可能与降低精氨酸酶、乙酰胆碱酯酶和腺苷脱氨酶的活性,上调NO水平,抑制PDE-5活性及影响细胞信号转导等机制相关。     

基于PI3K/AKT/eNOS信号通路探讨健脾利湿化瘀方抑制前列腺癌血管生成的实验研究

[目的]在健脾利湿化瘀方临床疗效和前期实验的基础上,继续探讨健脾利湿化瘀方及其拆方在抑制人前列腺癌PC-3细胞荷瘤小鼠血管生成方面的作用,及其对PI3K/AKT/eNOS信号通路的影响。[方法]选取6~8周雄性裸鼠36只建立人前列腺癌荷瘤小鼠模型,随机分为空白对照组、西药组、全方组、君药组、臣药组和佐药组。实验各组干预14 d后处死取血称瘤质量,计算抑瘤率;使用酶联免疫吸附实验(ELISA)检测血清中血管内皮生长因子(VEGF)、内皮型一氧化氮合成酶(eNOS);免疫组化法观察各组瘤组织中VEGF、ANG1的表达;蛋白免疫印迹法(Western Blot)检测肿瘤组织中磷脂酰肌3-羟激酶(PI3K)、丝氨酸-苏氨酸蛋白激酶(AKT)、磷酸化的AKT(p-AKT)、eNOS蛋白表达水平。[结果]各给药组瘤体积、瘤质量均低于空白对照组(P<0.01),西药组、全方组、君药组、臣药组、佐药组的抑瘤率分别为53.95%、41.05%、19.21%、27.37%、31.32%,拆方组间比较,佐药组抑瘤效果较好,但无统计学差异(P>0.05);ELISA检测显示:各组VEGF、eNOS含量均低于空白对照组,其中西药组、全方组显著降低(P<0.05或P<0.01);Western Blot结果显示:各组PI3K、eNOS的蛋白表达均显著降低(P<0.05或P<0.01),AKT蛋白表达无显著变化(P>0.05),p-AKT呈降低趋势(P<0.05),其中西药组、全方组、佐药组呈显著下降(P<0.01)。[结论]尽管与西药对比抑瘤作用较弱,但中药健脾利湿化瘀方对前列腺癌在血管生成方面具有确切的抑制作用,尤其是具有化瘀消癥散结作用的佐药组,可能与姜黄、王不留行中某些有效成分能够下调PI3K、eNOS蛋白,影响AKT磷酸化,阻断PI3K/AKT/eNOS信号通路相关。     

豨莶草提取物对冠状动脉结扎大鼠心肌AKT及ERK1/2表达的影响

[目的] 通过豨莶草提取物对心肌缺血大鼠模型心肌蛋白激酶（AKT）及丝裂原活化蛋白激酶1/2（ERK1/2）表达的影响,探讨其对大鼠心肌缺血性损伤的保护作用。[方法] 将90只SD大鼠雄性随机分为6组,每组15只,分别为假手术组、模型对照组、豨莶草提取物低剂量组、豨莶草提取物中剂量组、豨莶草提取物高剂量组、硝酸异山梨酯组。构建大鼠心肌缺血损伤模型,用无创缝合线在左心耳根部下方2 mm处结扎。连续给药4周后。大鼠腹主动脉采血检测心肌酶,采用苏木精-伊红（HE）染色法观察缺血区心肌损伤级别,采用免疫组化法检测心肌细胞AKT及ERK1/2的表达。[结果] 与模型对照组相比较,豨莶草提取物低剂量组、中剂量组和高剂量组血清谷草转氨酶（AST）显著降低;豨莶草提取物低剂量组、中剂量组和高剂量组血清乳酸脱氢酶（LDH）有所降低,但未见显著性差异;豨莶草提取物高剂量组血清肌酸激酶（CK）显著降低;豨莶草提取物中剂量组、高剂量组血清肌酸磷酸激酶（CK-MB）显著降低;豨莶草提取物低剂量组、中剂量组、高剂量组缺血区心肌组织HE染色分级显著降低;豨莶草提取物低剂量组、中剂量组、高剂量组缺血区心肌细胞AKT的表达显著增加;豨莶草提取物中剂量组、高剂量组缺血区心肌细胞ERK1/2表达显著增加。[结论] 豨莶草提取物可通过增加心肌缺血区AKT、ERK1/2表达而产生对心肌损伤的保护作用。