Investigation of apoptotic and antiproliferative effects of Turkish natural tetraploids Trifolium pratense L. extract on C6 glioblastoma cells via light and electron microscopy

ABSTRACT

Glioblastoma (GBM) is the most common type of primary brain tumors in adults, characterized by its ability to proliferate rapidly and its tendency to aggressively and strongly invaded the surrounding brain tissue. The standard treatment approach of GBM is surgical resection followed by simultaneous chemotherapy and radiation. However, a significant number of GBM cases develop resistance to currently used chemotherapeutic drugs. Therefore, there is a need for the development of new chemotherapeutic agents. Trifoliumpratense L. is an endemic plant containing various isoflavones such as biochanin A, genistein, daidzein, and formononetin in high concentrations, and it has been shown in various studies that these molecules can function as anticancer agents. The present study was designed to determine the effect of the possible anticarcinogenic effects of the Trifolium pratense L. which grown in our country and to obtain new treatment approaches alternative to the classical treatment protocols applied in the treatment of GBM. C6 glioblastoma cells were cultured with Trifolium pratense L. Cell proliferation, apoptotic cell morphology, and cell structure were evaluated with CCK8, Annexin V, cytochrome c, CD117, and Betatubulin labeling, respectively. And also, investigated effects of this Turkish tetraploid on GBM by TEM. Decreased cell proliferation and increased number of apoptotic cells were observed depending on the increasing doses of Trifolium pratense L. In addition, intense morphological changes were detected depending on increasing doses. In this context, we believe that the plant Trifolium pratense L., may be a new alternative and adjuvant agent for the treatment of GBM.

KEYWORDS:

• Glioblastoma
• Trifolium pratense L.
• apoptosis
• biochanin A
• genistein

Disclosure statement

No potential conflict of interest was reported by the authors.

Author contributions

Gamze Tanrıverdi designed the Project, evaluated histological results and drafted manuscript, Belisa Kaleci and Aynur Abdulova performed the experimental procedure and the immunohistochemical staining. Tuğba Ekiz-Yılmaz prepared the electron microscopic samples. Havva Atar and Hatice Çölgeçen grew the plant and prepared the extract. All authors contributed to the final version of the manuscript.

Additional information

Funding

This work was supported by the Scientific Research Projects Coordination Unit of Istanbul University-Cerrahpasa [TSA-2019-33277].