guanosine-triphosphate and Carcinoma--Non-Small-Cell-Lung

Lung cancer remains the leading cause of cancer-related deaths in men and women worldwide, and 85% of these patients have non-small cell lung cancer. In recent years, the clinical use of targeted drug therapy and immune checkpoint inhibitors has dramatically changed the treatment landscape for advanced NSCLC. The mechanism and the value of targeted therapies have been a hot topic of research, as. 近年来，靶向药物治疗和免疫检查点抑制剂的临床应用极大地改变了晚期非小细胞肺癌(NSCLC)的治疗格局。表皮生长因子受体和间变性淋巴瘤受体酪氨酸激酶等驱动基因改变NSCLC的TKI靶向治疗均已取得了良好临床疗效，而Kirsten大鼠肉瘤病毒基因同源物(KRAS)作为较早发现和突变频率较高的癌基因之一，其靶向药物治疗研究进展缓慢，法尼基转移酶抑制剂、KRAS信号通路下游蛋白抑制剂等靶向治疗研究均未取得预期结果，使得KRAS长期以来被定义为"不可成药的靶点"。KRAS蛋白作为分子开关，通过与三磷酸鸟苷结合而被激活，引发系列级联反应，在细胞增殖和有丝分裂中发挥作用。KRAS突变的NSCLC患者对内科系统性治疗反应性差，预后不佳。随着对KRAS晶体结构认识的不断深入，研究者发现了KRAS潜在的治疗位点，进而开发出了多个直接针对KRAS的靶向药物，尤其是KRAS G12C抑制剂，如AMG510(sotorasib)和MRTX849(adagrasib)，其临床试验获得了令人鼓舞的结果。文章在系统介绍KRAS突变NSCLC患者临床特征及内科治疗方法的基础上，重点就KRAS靶向治疗的研究进展进行了总结和展望。.

Topics: Carcinoma, Non-Small-Cell Lung; Female; Genes, ras; Guanosine Triphosphate; Humans; Immune Checkpoint Inhibitors; Lung Neoplasms; Male; Mutation; Proto-Oncogene Proteins p21(ras)

Topics: Animals; Antineoplastic Agents; Carcinoma, Non-Small-Cell Lung; Guanosine Triphosphate; Humans; Lung Neoplasms; Mice; Mutation; Proto-Oncogene Proteins p21(ras)

Rho GTPases play an essential role in many cellular processes, including cell cycle progress, cell motility, invasion, migration, and transformation. Several studies indicated that the dysregulation of Rho GTPase signaling is closely related to tumorigenesis. Rho GEFs considered being positive regulators of Rho GTPase, promoting the dissociation of Rho protein from GDP and binding to GTP, thus activating the downstream signaling pathway. Herein, we demonstrated that ARHGEF3, a member of the Rho GEFs family, played an important role in non-small cell lung cancer (NSCLC). We found that ARHGEF3 was highly expressed in non-small cell lung cancer and facilitated cancer cell proliferation of NSCLC cells in vitro and in vivo. Further studies demonstrated that ARHGEF3 enhanced the protein homeostasis of ATP-citrate lyase (ACLY) by reducing its acetylation on Lys17 and Lys86, leading to the dissociation between ACLY and its E3 ligase-NEDD4. Interestingly, this function of ARHGEF3 on the protein homeostasis of ACLY was independent of its GEF activity. Taken together, our findings uncover a novel function of ARHGEF3, suggesting that ARHGEF3 is a promising therapeutic target in non-small cell lung cancer.

Topics: Adenosine Triphosphate; ATP Citrate (pro-S)-Lyase; Carcinoma, Non-Small-Cell Lung; Cell Proliferation; Guanosine Triphosphate; Humans; Lung Neoplasms; rho GTP-Binding Proteins; Rho Guanine Nucleotide Exchange Factors; Ubiquitin-Protein Ligases

Ginsenosides have previously been demonstrated to effectively inhibit cancer cell growth and survival in both animal models and cell lines. However, the specific ginsenoside component that is the active ingredient for cancer treatment through interaction with a target protein remains unknown. By an integrated quantitative proteomics approach via affinity mass spectrum (MS) technology, we deciphered the core structure of the ginsenoside active ingredient derived from crude extracts of ginsenosides and progressed toward identifying the target protein that mediates its anticancer activity. The Tandem Mass Tag (TMT) labeling quantitative proteomics technique acquired 55620 MS/MS spectra that identified 5499 proteins and 3045 modified proteins. Of these identified proteins, 224 differentially expressed proteins and modified proteins were significantly altered in nonsmall cell lung cancer cell lines. Bioinformatics tools for comprehensive analysis revealed that the Ras protein played a general regulatory role in many functional pathways and was probably the direct target protein of a compound in ginsenosides. Then, affinity MS screening based on the Ras protein identified 20(s)-protopanaxadiol, 20(s)-Ginsenoside Rh2, and 20(s)-Ginsenoside Rg3 had affinity with Ras protein under different conditions. In particular, 20(s)-protopanaxadiol, whose derivatives are the reported antitumor compounds 20(s)-Ginsenoside Rh2 and 20(s)-Ginsenoside Rg3 that have a higher affinity for Ras via a low KD of 1.22 μM and the mutation sites of G12 and G60, was demonstrated to play a core role in those interactions. Moreover, the molecular mechanism and bioactivity assessment results confirmed the identity of the chemical ligand that was directly acting on the GTP binding pocket of Ras and shown to be effective in cancer cell bioactivity profiles.

Topics: Animals; Antineoplastic Agents, Phytogenic; Carcinoma, Non-Small-Cell Lung; Cell Cycle; Cell Line, Tumor; Cell Survival; Ginsenosides; Guanosine Triphosphate; Humans; Lung Neoplasms; Molecular Docking Simulation; Neoplasm Proteins; Protein Binding; Protein Conformation; Proteomics; ras Proteins; Sapogenins

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death in the world. Inhibitor of differentiation 1 (Id1) is overexpressed in NSCLC and involved in promoting its progression and metastasis. Identifying natural compounds targeting Id1 may have utility in NSCLC treatment. Here, we sought to determine whether the anti-tumor activities of

Topics: A549 Cells; alpha7 Nicotinic Acetylcholine Receptor; Animals; Antineoplastic Agents; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Flavanones; Flavonoids; Guanosine Triphosphate; Humans; Inhibitor of Differentiation Protein 1; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Neoplasm Metastasis; Phenotype; Phosphorylation; Plant Extracts; Scutellaria; Shelterin Complex; Telomere-Binding Proteins

RAS mutations are frequent in human cancer, especially in pancreatic, colorectal and non-small-cell lung cancers (NSCLCs)

Topics: Animals; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Proliferation; Cellular Senescence; Guanosine Triphosphate; Humans; Lung Neoplasms; Mice; Mitogen-Activated Protein Kinase Kinases; Mutation; Protein Kinase Inhibitors; Protein Tyrosine Phosphatase, Non-Receptor Type 11; Proto-Oncogene Proteins p21(ras); Xenograft Model Antitumor Assays

Protein kinase C (PKC) ε, a key signaling transducer implicated in mitogenesis, survival, and cancer progression, is overexpressed in human primary non-small cell lung cancer (NSCLC). The role of PKCε in lung cancer metastasis has not yet been established.. Here we show that RNAi-mediated knockdown of PKCε in H358, H1299, H322, and A549 NSCLC impairs activation of the small GTPase Rac1 in response to phorbol 12-myristate 13-acetate (PMA), serum, or epidermal growth factor (EGF). PKCε depletion markedly impaired the ability of NSCLC cells to form membrane ruffles and migrate. Similar results were observed by pharmacological inhibition of PKCε with εV1-2, a specific PKCε inhibitor. PKCε was also required for invasiveness of NSCLC cells and modulated the secretion of extracellular matrix proteases and protease inhibitors. Finally, we found that PKCε-depleted NSCLC cells fail to disseminate to lungs in a mouse model of metastasis.. Our results implicate PKCε as a key mediator of Rac signaling and motility of lung cancer cells, highlighting its potential as a therapeutic target.

Topics: Animals; Carcinoma; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Disease Progression; Enzyme Activation; Extracellular Matrix; Gene Expression Regulation, Neoplastic; Guanosine Triphosphate; Humans; Lung Neoplasms; Male; Mice; Mice, Nude; Neoplasm Metastasis; Protein Kinase C-epsilon; rac GTP-Binding Proteins; RNA Interference; Signal Transduction

The cancer stem cell (CSC) theory predicts that a small fraction of cancer cells possess unique self-renewal activity and mediate tumor initiation and propagation. However, the molecular mechanisms involved in CSC regulation remains unclear, impinging on effective targeting of CSCs in cancer therapy. Here we have investigated the hypothesis that Rac1, a Rho GTPase implicated in cancer cell proliferation and invasion, is critical for tumor initiation and metastasis of human non-small cell lung adenocarcinoma (NSCLA). Rac1 knockdown by shRNA suppressed the tumorigenic activities of human NSCLA cell lines and primary patient NSCLA specimens, including effects on invasion, proliferation, anchorage-independent growth, sphere formation and lung colonization. Isolated side population (SP) cells representing putative CSCs from human NSCLA cells contained elevated levels of Rac1-GTP, enhanced in vitro migration, invasion, increased in vivo tumor initiating and lung colonizing activities in xenografted mice. However, CSC activity was also detected within the non-SP population, suggesting the importance of therapeutic targeting of all cells within a tumor. Further, pharmacological or shRNA targeting of Rac1 inhibited the tumorigenic activities of both SP and non-SP NSCLA cells. These studies indicate that Rac1 represents a useful target in NSCLA, and its blockade may have therapeutic value in suppressing CSC proliferation and metastasis.

Topics: Adenocarcinoma; Animals; Carcinoma, Non-Small-Cell Lung; Cell Adhesion; Cell Line, Tumor; Cell Movement; Cell Proliferation; Gene Knockdown Techniques; Guanosine Triphosphate; Humans; Lung; Lung Neoplasms; Mice; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplastic Stem Cells; rac1 GTP-Binding Protein; RNA, Small Interfering; Side-Population Cells

Expression of the tumor suppressor deleted in liver cancer-1 (DLC-1) is lost in non-small cell lung (NSCLC) and other human carcinomas, and ectopic DLC-1 expression dramatically reduces proliferation and tumorigenicity. DLC-1 is a multi-domain protein that includes a Rho GTPase activating protein (RhoGAP) domain which has been hypothesized to be the basis of its tumor suppressive actions. To address the importance of the RhoGAP function of DLC-1 in tumor suppression, we performed biochemical and biological studies evaluating DLC-1 in NSCLC. Full-length DLC-1 exhibited strong GAP activity for RhoA as well as RhoB and RhoC, but only very limited activity for Cdc42 in vitro. In contrast, the isolated RhoGAP domain showed 5- to 20-fold enhanced activity for RhoA, RhoB, RhoC, and Cdc42. DLC-1 protein expression was absent in six of nine NSCLC cell lines. Restoration of DLC-1 expression in DLC-1-deficient NSCLC cell lines reduced RhoA activity, and experiments with a RhoA biosensor demonstrated that DLC-1 dramatically reduces RhoA activity at the leading edge of cellular protrusions. Furthermore, DLC-1 expression in NSCLC cell lines impaired both anchorage-dependent and -independent growth, as well as invasion in vitro. Surprisingly, we found that the anti-tumor activity of DLC-1 was due to both RhoGAP-dependent and -independent activities. Unlike the rat homologue p122RhoGAP, DLC-1 was not capable of activating the phospholipid hydrolysis activity of phospholipase C-delta1. Combined, these studies provide information on the mechanism of DLC-1 function and regulation, and further support the role of DLC-1 tumor suppression in NSCLC.

Topics: Carcinoma, Non-Small-Cell Lung; Cell Movement; Collagen; DNA Primers; Drug Combinations; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; GTPase-Activating Proteins; Guanosine Triphosphate; Humans; Hydrolysis; Laminin; Lung Neoplasms; Neoplasm Invasiveness; Phospholipase C delta; Polymerase Chain Reaction; Proteoglycans; rho GTP-Binding Proteins; rhoA GTP-Binding Protein; rhoB GTP-Binding Protein; rhoC GTP-Binding Protein; Tumor Cells, Cultured; Tumor Stem Cell Assay; Tumor Suppressor Proteins