Tumor Cell Autophagy Signaling Analysis

Autophagy, a cellular degradation process, plays a dual role in cancer by promoting survival under stress and enabling tumor progression when dysregulated. At Alfa Cytology, we offer a suite of advanced analytical services to dissect the autophagy signaling pathways in tumor cells.

Introduction

Autophagy is a lysosomal catabolic process involving the degradation of cellular materials, protein aggregates and dysfunctional organelles to maintain cellular homeostasis. Strikingly, autophagy exhibits a dual role in cancer. On the one hand, it promotes the clearance of transformed cells and suppresses tumorigenesis, while cytoprotective autophagy plays a role in the maintenance of cancer.

It has now been shown that: autophagic signaling pathways support cancer growth and progression in an oxidative and hypoxic environment adapted to TME; autophagy contributes to the regulation of metabolic switches to produce sufficient levels of high-energy metabolites, including amino acids, ketones, glutamine, and free fatty acids for cancer cell survival; autophagy has a key role in the regulation of tumor-associated fibroblasts, leading to different cytokines and paracrine autophagy promotes immune evasion to suppress antitumor immunity; autophagy inhibitors enhance response to immunotherapy through infiltration of immune cells into TME ecological niches; autophagy in TME maintains and supports cancer stem cell survival, leading to chemoresistance and treatment relapse.

Our Services

A continuing challenge among researchers of autophagic processes and cancer growth and treatment is that the transduction and regulation of autophagic signaling in the tumor microenvironment (TME) is not well understood, and therefore many of these studies lack the support of molecular mechanistic theories. Alfa Cytology has established an innovative tumor microenvironment center technology platform and is developing multiple technologies to provide high quality solutions and services to our global collaborators. We believe that with the in-depth study of autophagy signal transduction and autophagy regulation mechanisms in the tumor microenvironment,, we will soon elucidate those unknown puzzles. Thus, autophagy can be used as a target for tumor action, and tumor prevention and control can be achieved by regulating autophagy.

Key Technologies

Morphological changes in different stages of autophagy were directly observed under transmission electron microscopy, since autophagosomes belong to subcellular structures. In the early stage, the main feature is a crescent-shaped, single-layer or multi-layer membrane; in the middle stage, the double-layer or multi-layer vacuole-like structure is the autophagosome structure, which contains cytoplasmic components, such as mitochondria and endoplasmic reticulum; in the later stage, autophagolysosomes are formed, monolayer membranes, and cytoplasmic components have been degraded.

Indirect observation of RFP-GFP-LC3 double fluorescent labeling. When autophagosome and lysosome are not fused, both RFP fluorescence and GFP fluorescence exist. When autophagosome and lysosome are fused, GFP is quenched, and only red fluorescence can be observed.

Indirect observation of GFP-LC3 single fluorescent labeling. Using the principle that LC3 aggregates during autophagy formation, that is, in the absence of autophagy, the GFP-LC3 fusion protein diffuses in the cytoplasm.

MDC staining. MDC is an eosinophilic fluorescent pigment, and it is non-specific staining because it can stain all acidic vesicles. Autophagosomes are acidic vesicles that can be colored.

Research on autophagy-related proteins. Western blot detection of LC3 ll /l, p62, Beclin-1, ULK1 protein expression.

Alfa Cytology is committed to supporting scientists in making breakthrough scientific discoveries and developing new applications to accelerate new drug discovery and scientific diagnosis and treatment. Our high-performance scientific instruments and high-value solutions enable scientists to explore the mysteries of life at the tumor microenvironment level. Please tell us your project requirements, and we will provide you with a full service from solution to report. If you have any questions, please feel free to contact us.

! For research use only.