Camptothecin (CPT) has strong antitumor activity and is used as an anticancer therapeutic agent. To better understand and decipher the pathway of CPT biosynthesis in Camptotheca acuminata, the main purpose here was focused on creating an effective extraction strategy for a rich intermediate metabolite profile. In the present study, a 70% aqueous acetonitrile was verified as an optimal extraction solvent for microwave-assisted extraction (MAE) of metabolites by spiking experiments. Based on multi-objective optimization, the best extraction conditions of a solid-liquid ratio of 1:20, microwave power of 230 W, and a time of 4 min were achieved using a full factorial 34 experimental design. Crude extracts obtained from the shoot apex of C. acuminata using MAE have been qualitatively profiled by high-performance liquid chromatography coupled with linear ion trap quadrupole-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap-MS/MS) and a HPLC triple quadrupole-MS (HPLC-TSQ-MS) analysis was conducted for their metabolite content in different tissues. CPT, and ten related metabolites and their isomers, including tryptamine, loganic acid, secologanic acid, strictosidinic acid, strictosamide, strictosamide epoxide, strictosamide diol, strictosamide ketolactam, pumiloside, and deoxypumiloside, were detected and tentatively identified. Scanning electron microscopy (SEM) imaging of the shoot apex demonstrated that severe cell disruption was evident after intensified extraction processes. The study showed the difference of metabolite profiles and the enhancement of metabolite content after microwave-pretreated techniques, and the established MAE procedure is an effective methodology to preserve valuable metabolite compounds for analysis.
BackgroundLung cancer is a malignant lung tumor characterized by uncontrolled cell growth in tissues of the lung. Myeloid leukemia factor 1 (MLF1) is a protein involved in myeloid cell differentiation which regulates the cell cycle and the expression of numerous genes. The role of MLF1 in hematologic cancers is well established, As yet, there is little knowledge of MLF1 expression and biological function in lung cancer. Here, we investigated the role of MLF1 in lung cancer using a variety of cell lines along with patient samples to determine whether MLF1 played a significant role in this devastating disease.Materials and MethodsLung cancer cell lines (A549, H1975, HCC827, and NCI‐H460) and primary lung tissue were used to assess the relative levels of MLF1 in lung cancer. The lung cancer cell line A549 was lentivirally transfected to silence endogenous MLF1 gene expression, which was confirmed by real‐time quantitative PCR (qPCR). Cell proliferation was assessed through fluorescence imaging and MTT assays. Cell cycle analysis was performed utilizing flow cytometry and formation of cell colonies evaluated microscopically.ResultsProliferation of A549 cells was significantly inhibited in cells where MLF1 was silenced compared to controls. Cell cycle analysis indicated that cell cycle phases were not significantly changed upon silencing of MLF1 in lung cancer cells. A significant increase in apoptosis was observed in MLF1‐knockdown cells, while a significant decrease in number of cell colonies formed was observed in MLF1‐knockdown cells compared to controls. In most, but not all, human lung cancer tissue samples MLF1 was upregulated.ConclusionIn summary, MLF1 promotes the proliferation and colony forming abilities of lung cancer cells and significantly decreases apoptosis while have no impact on cell cycle. Further studies with larger sample sizes are needed 1) to conclude whether human lung cancers upregulate MLF1, 2) to reveal the mechanism of action for MLF1 in lung carcinogenesis and 3) investigate MLF1 gene therapy for the treatment of lung cancer.Support or Funding InformationThis work was supported by Key Program of Natural Science Research of Higher Education of Anhui Province (Grant KJ2016A472 KJ2017A241), Key program for excellent young talents in college and university of Anhui province (Grant gxyqZD2016168), National Natural Science Foundation of China (Grant 81772493), and College students' innovation and entrepreneurship training project (Grant 201610367007)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.