Calcitriol potentially alters HeLa cell viability via inhibition of autophagy Setiawan I, Lesmana R, Goenawan H, Suardi D, Gatera VA, Abdulah R, Judistiani RT, Supratman U, Setiabudiawan B,

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Calcitriol potentially alters HeLa cell viability via inhibition of autophagy Iwan Setiawan¹, Ronny Lesmana¹, Hanna Goenawan², Dodi Suardi³, Vesara Ardhe Gatera⁴, Rizky Abdulah⁴, Raden Tina Dewi Judistiani⁵, Unang Supratman⁶, Budi Setiabudiawan⁷ ¹ Departement of Biomedical Science, Physiology Division, Faculty of Medicine; Physiology Molecular, Biological Activity Division, Central Laboratory; Vitamin D Centre, Faculty of Medicine, Universitas Padjadjaran, Bandung, West Java, Indonesia ² Departement of Biomedical Science, Physiology Division, Faculty of Medicine; Physiology Molecular, Biological Activity Division, Central Laboratory, Universitas Padjadjaran, Bandung, West Java, Indonesia ³ Departement of Obstetrics and Gynecology, Division of Oncology and Gynecology, Faculty of Medicine, Hasan Sadikin Hospital - Universitas Padjadjaran, Bandung, West Java, Indonesia ⁴ Departement of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Universitas Padjadjaran, Bandung, West Java, Indonesia ⁵ Vitamin D Centre; Departement of Public Health, Faculty of Medicine, Universitas Padjadjaran, Bandung, West Java, Indonesia ⁶ Physiology Molecular, Biological Activity Division, Central Laboratory; Departement of Chemistry, Faculty of Mathematics and Natural Science, Universitas Padjadjaran, Bandung, West Java, Indonesia ⁷ Vitamin D Centre; Departement of Child Health, Faculty of Medicine, Universitas Padjadjaran, Bandung, West Java, Indonesia Correspondence Address: Ronny Lesmana, Departement of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Bandung, West Java, 45363; Physiology Molecular, Biological Activity Division, Central laboratory, Bandung, West Java, Indonesia, 45363. Vitamin D Centre, Faculty of Medicine, Bandung, West Java, 45363 Indonesia Source of Support: None, Conflict of Interest: None DOI: 10.4103/jcrt.JCRT_82_20

Objective: This study was conducted to evaluate the effect of Calcitriol on cellular death in HeLa cells via autophagy and turn over due to mitochondria homeostasis. Methods: HeLa cell lines were grown in 24-well plates and treated with Calcitriol at varying doses (0.013 μM-0.325 μM) for varying time periods (2, 6, 12, and 18 h). Cell proteins were extracted with scrapers and lysed using RIPA buffer. Western blots were performed for proteins involved with autophagy (Lc3, p62), signaling (mTOR, PI3K, HIF1α), mitochondria (PGC1α, COX4, and Tom 20), and apoptosis (Caspase 3, Caspase 9, and PARP). Protein carbonyl levels were determined by measuring the indirect ROS level. An inhibition study using L-mimosine was performed to analyze the significance of HIF1α. Results: Calcitriol treatment induced cytotoxicity in a dose- and time-dependent manner and caused growth arrest in HeLa cells. The PI3K-AKT-mTOR pathway was activated, leading to inhibition of autophagy and alterations in mitochondria biogenesis homeostasis. Treatment with Calcitriol produced protein carbonyl levels similar to those in the cisplatin-treated and control groups. Increased ROS levels may cause toxicity and induce cell death specifically in cancer cells but not in normal cells. The inhibition of HIF1α partially rescued the HeLa cells from the toxic effects of Calcitriol treatment. Conclusion: We suggest that Calcitriol may shut down mitochondrial homeostasis in HeLa cells by inducing the PI3K-AKT-mTOR pathway and inhibiting autophagy, which leads to cell death.

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    -  Setiawan I
    -  Lesmana R
    -  Goenawan H
    -  Suardi D
    -  Gatera VA
    -  Abdulah R
    -  Judistiani RT
    -  Supratman U
    -  Setiabudiawan B

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