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Generation of reporter cell lines for factors inducing muscle wasting in cancer cachexia.

Analytical biochemistry (2020-08-02)
Zhipeng Cao, Irvin Jose, Jason Glab, Hamsa Puthalakath, Laura D Osellame, Nick J Hoogenraad

Rapidly identifying cachexia-inducing factors that directly induce muscle wasting is an existing challenge. We developed two reporter cell lines that allow swift detection of such factors in blood from patients. C2C12 myoblasts were used for the establishment of reporter cells. A luciferase reporter gene, driven by promoters of wasting genes, Muscle RING-finger protein-1 (MuRF1) and Muscle Atrophy F-Box Protein (MAFbx/Atrogin-1) were used for the construction of reporter constructs. Increased expression of these genes in muscle tissue under wasting conditions was shown in vivo and in vitro. We found these reporter cell lines could detect factors associated with cancer cachexia, such as myostatin (Mstn), activin A, and TNF-α. We further investigated the capacity to directly detect a cachectic state using plasma samples from cachectic mice and cancer patients. Activation of the reporter cell lines was observed by the addition of plasma from mice with cancer cachexia and serum samples from patients with pancreatic or colorectal cancer. These results indicate that the reporter cell lines are competent as a tool for screening cachexia-inducing factors and potentially distinguishing a cachectic state induced by cancer.

Product Number
Product Description

Horse Serum, Donor herd, USA origin, sterile-filtered, suitable for cell culture, suitable for hybridoma
Hexadimethrine bromide, ≥95%
Puromycin dihydrochloride from Streptomyces alboniger, ≥98% (HPLC), powder