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

Aleksandar Janev,Žiga Sardoč,Taja Železnik Ramuta,Larisa Tratnjek,Mateja Erdani Kreft,Tomaž Smrkolj,Slavko Mojsilović,Milena Taskovska,Hristina Obradović

doi:10.3389/fbioe.2021.690358

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.

Highlights

The incidence of bladder cancer is steadily increasing, especially in industrialized countries (Bray et al, 2018)
The confluent cultures of T24, RT4 normal porcine urothelial (NPU), MCF7, and MCF10a were treated with an appropriate culture medium and human amniotic membrane (hAM) homogenate for 24 h (Figures 2A–J)
We found that hAM homogenate caused the detachment of several cancer cell lines

Summary

Introduction

The incidence of bladder cancer is steadily increasing, especially in industrialized countries (Bray et al, 2018). The predominant histologic type of bladder cancer is urothelial carcinoma and out of all newly diagnosed cases, approximately 75% are non-muscle invasive bladder cancer (NMIBC) and 25% are muscle-invasive bladder cancer (MIBC) (Sanli et al, 2017; Patel et al, 2020). The main challenge of NMIBC management is its high recurrence rate. Approximately 50–70% of NMIBC cases will recur and 15–20% will progress to MIBC (Sylvester et al, 2006; Isharwal and Konety, 2015; Knowles and Hurst, 2015; Patel et al, 2020). Due to the high recurrence rate and consequent disease monitoring requirements, bladder cancer has one of the highest lifetime treatment costs per patient among cancers (Sievert et al, 2009). The prognosis for patients with NMIBC is very encouraging as the 5-year survival rate is approximately 90%, the 5-year survival rate of patients with the metastatic disease is still merely 6% (Funt and Rosenberg, 2017)

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