Detrimental Effect of Various Preparations of the Human Amniotic Membrane Homogenate on the 2D and 3D Bladder Cancer In vitro Models

Abstract

Despite being among the ten most common cancers with high recurrence rates worldwide, there have been no major breakthroughs in the standard treatment options for bladder cancer in recent years. The use of a human amniotic membrane (hAM) to treat cancer is one of the promising ideas that have emerged in recent years. This study aimed to investigate the anticancer activity of hAM homogenate on 2D and 3D cancer models. We evaluated the effects of hAM homogenates on the human muscle invasive bladder cancer urothelial (T24) cells, papillary cancer urothelial (RT4) cells and normal porcine urothelial (NPU) cells as well as on human mammary gland non-tumorigenic (MCF10a) cells and low-metastatic breast cancer (MCF7) cells. After 24 h, we observed a gradual detachment of cancerous cells from the culture surface, while the hAM homogenate did not affect the normal cells. The most pronounced effect hAM homogenate had on bladder cancer cells; however, the potency of their detachment was dependent on the treatment protocol and the preparation of hAM homogenate. We demonstrated that hAM homogenate significantly decreased the adhesion, growth, and proliferation of human bladder invasive and papillary cancer urothelial cells and did not affect normal urothelial cells even in 7-day treatment. By using light and electron microscopy we showed that hAM homogenate disrupted the architecture of 2D and 3D bladder cancer models. The information provided by our study highlights the detrimental effect of hAM homogenate on bladder cancer cells and strengthens the idea of the potential clinical application of hAM for bladder cancer treatment.

Human amniotic membrane inhibits migration and invasion of muscle-invasive bladder cancer urothelial cells by downregulating the FAK/PI3K/Akt/mTOR signalling pathway

“…In addition to the effect on proliferation, hAM scaffolds also reduce the invasive potential and the expression of mesenchymal markers (Snail, Slug, N-cadherin) in cancer urothelial cells 18 . Furthermore, we were the first to demonstrate that the hAM homogenate decreased the adhesion of cancer urothelial cells, impeded their growth dynamics and disrupted the 3D structure of bladder cancer spheroids, while it had no such deleterious effects on normal urothelial cells 22 .…”

Antiproliferative and apoptotic effects of conditioned medium released from human amniotic epithelial stem cells on breast and cervical cancer cells

“… 13 A recent study from Janev et al demonstrated that hAM homogenate significantly decreased the adhesion, growth, and proliferation of human bladder invasive and papillary cancer urothelial cells, but did not affect normal urothelial cells. 26 Besides, Protein extracts obtained from processed hAM have been shown to suppress metabolic activity in hepatocarcinoma cell lines, but had no effect on metabolic activity inhibition or protein and DNA content in non-tumorigenic cell lines.…”

“…Although the precise factors and mechanisms responsible for the anti-cancer roles of amniotic membrane remain unknown, previous studies have hypothesized biological mechanisms, including down-regulation of cancer cells’ gene expression associated with cell cycle progression, inhibition of heat shock protein 90, and secretion of some mediators, such as interleukin (IL)-2, IL-4, IL-8, IL-10, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TNF-β), along with indoleamine 2,3-dioxygenase (IDO), prostaglandin E2 (PGE2), and HLA-G. 6 , 9 , 26 , 27 Kang and colleagues established that hAECs via expression of cytotoxic factors, such as TNF-α, TGF-ß, or ILs, reduced the amount of growth and proliferation of MDA-MB-231 cancer cells. 9 A plenty of studies shown the link between inflammation and development of cancer.…”

Methods and criteria for validating the multimodal functions of perinatal derivatives when used in oncological and antimicrobial applications

“…The investigation of DNA synthesis analyzes the effects of PnDs on tumor cell proliferation. Incorporation of radiolabeled DNA precursor 3H-thymidine ( Ayuzawa et al, 2009 ; Magatti et al, 2012 ; Marleau et al, 2012 ; Yuan et al, 2013 ; Riedel et al, 2019 ; Ramuta et al, 2020b ) into new strands of chromosomal DNA or more recent analogs, BrdU (bromoeoxyuridin) ( Tian et al, 2010 ; Gauthaman et al, 2012 ; Han et al, 2014 ; Hendijani et al, 2015 ) and EdU (5-ethynyl-2 deoxyuridine) ( Wang M et al, 2015 ; Janev et al, 2021 ), have been widely used to assess de novo DNA synthesis. 3H-thymidine detection requires radioactive labeling followed by detection with a scintillation beta-counter, while BrdU can be easily detected by antibodies followed by flow cytometry or by immunohistochemistry.…”