Anti-cancer Assays for Next-generation Probiotics

Creative Biolabs, a world-leading CRO, is a recognized expert in the services of live biotherapeutics. We are a passionate team of scientists and we have the expertise and capability to deliver high-quality next-generation probiotics (NGPs) solutions.

Gut Microbiota and Cancer

The gut microbiota and its relationship to cancer development/treatment have been extensively studied over the past few years. Probiotics can increase and decrease the production of anti-inflammatory cytokines, which play an important role in preventing carcinogenesis. They also play an important role in activating phagocytes to abolish early-stage cancer cells. Probiotics, including Lactobacillus, have been shown to have tumor suppressive effects in colorectal cancer cell lines and mouse tumor models. Currently, there is a growing interest in NGPs as therapeutic agents that alter gut microbiota and impact cancer development. F. prausnitzii, A. muciniphila, B. fragilis and belong to the NGPs group and can be useful in cancer patients through several mechanisms.

Mechanisms of Probiotic's Anti-cancer

  • Antiproliferative activity, according to many studies, probiotics and post-biological agents have shown antiproliferative activity on cancer cell lines.
  • Induction of cancer cell apoptosis, or genetically programmed cell death, is a mechanism used to control cell numbers in multicellular organisms.
  • Produces compounds with cytoprotective (anti-proliferation and pro-apoptotic) effects on intestinal epithelial cells, such as organic acids, including short-chain fatty acids (SCFAs) and lactic acid (LA).

Mechanism of probiotics and prebiotics in cancer prevention.Fig.1 Mechanism of probiotics and prebiotics in cancer prevention. (Sharma, 2021)

Detection of Anti-cancer Activity at Creative Biolabs

Bacterial cell-free culture supernatants of probiotics trains have also been tested for preventing colonic cancer cell invasion in vitro. Strains showing in vitro inhibitory activity on tumor cell proliferation, induction of apoptotic cell death, and ability of cellular sphingolipids metabolism, have been recognized as promising candidates for cancer prevention.

  • Cytotoxicity Test Versus Normal Peripheral Blood Mononuclear Cells (PBMCs)

Peripheral blood is the accessible resource of human normal cells which can be used to assess the toxicity of extracts. About 1×105 mononucleate cells were injected into every single well of the 96-well microtiter plate. The cells were given two treatments: one with and one without serial dilutions of extracts.

  • Cytotoxicity Assay Against Cancerous Cell Lines

For the determination of the anticancer activity of the tested LAB supernatants, four different types of human cancer cell lines were used, and their growth inhibitory effects were assessed using an MTT assay.

  • Flow Cytometry Assessment

The anti-cancer mechanism of the most active anti-cancer lactic acid bacteria supernatant was examined by staining apoptotic and necrotic cells with dual nuclear fluorescent dyes (annexin V and PI), respectively.

In vitro toxicity assays.Fig.2 In vitro toxicity assays. (Soltani, 2021)

For more than a decade, Creative Biolabs has earned a global reputation for providing professional, tailored customer service and outstanding customer assistance to numerous clients. Our goal is to provide our customers with affordable development services and reliable results. Please feel free to contact us for anti-cancer activity assays of next-generation probiotics.


  1. Sharma, V.; et al. Probiotics and prebiotics having broad spectrum anticancer therapeutic potential: Recent trends and future perspectives. Current Pharmacology Reports. 2021, 7(2): 67-79.
  2. Soltani, S.; et al. Bacteriocins as a new generation of antimicrobials: toxicity aspects and regulations. FEMS Microbiol Rev. 2021, 45.

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