The Effect of Melatonin and Cisplatin Combination Using Copper-Transporting ATPase-1, P-Glycoprotein, and Gamma-Glutamylcysteinylglycine on Ovarian Cancer Biological Cell SKOV3
Authors: Adella Cut Adeya , Siregar M. Fidel Ganis , Putra Imam Budi , Hasibuan Poppy Anjelisa , Andrijono Andrijono , Bachtiar Adang , Lumbanraja Sarma N. , Nasution Iqbal Pahlevi 
Published: June 2022

Background: Ovarian cancer is fifth most common female cancer and third most common  cancer in Indonesia, but most are advanced stage patients that experiencing recurrence, which indicates resistance to treatment especially to cisplatin. Melatonin appears as an alternative that can support apoptotic effect of cisplatin as a chemotherapy regimen.
Aim: To determine effect of combination melatonin and cisplatin compared with cisplatin only chemotherapy on chemotherapy resistance with Copper Transporting ATPase-1 (CTR-1), P-glycoprotein (P-Gp), and Gamma-Glutamylcysteinylglycine (GSH)  biomarkers in ovarian cancer biological cells SKOV3
Methods: This research design was experimental laboratory, post-test only control group design, using SKOV3 cell culture. This study was performed in the SCTE IMERI FKUI laboratory and Integrated Laboratory FKUI. MTS assay was used to calculate IC50 of each materials. The materials used were melatonin (concentration was 25,50,100,200,300 nM), cisplatin (concentration was 0.1, 0.5, 1, 2, 5 mM), and doxorubicin (concentration 10,20,40,50,80,100,200 µM). IC50 melatonin was 1,841 mM, IC50 cisplatin was 117,5 µM, and IC50 doxorubicin was 14,72 µM. Samples were control negative group, IC50 doxorubicin as a control positive, IC50 cisplatin, IC50 melatonin, combination group of melatonin and cisplatin were 1xIC50, ¾xIC50, ½xIC50, and ¼xIC50. ANOVA and Bonferroni test were used for statistical test.
Results: Based on data processing, IC50 of melatonin was 1,841 mM, IC50 of doxorubicin was 14,72 mM, while IC50 of cisplatin was 117.5 μM. The mean expression of CTR-1 in IC50 melatonin group was 15.77 ± 0.21 and in IC50 cisplatin group was 10.87 ± 0.91, mean expression in IC50 doxorubicin group was 30,33 ±  0,4. Meanwhile, mean expression of CTR-1 in IC50 cisplatin was 7,37±0,7, and in combination 1 group (1xIC50 melatonin and 1xIC50 cisplatin) was 19,73±1.0,49. For P glycoprotein, mean expression in IC50 cisplatin was 16±1,59, in IC50 melatonin group was 7,37±0,21, in IC50 doxorubicin was 0, and in combination 1 group (1xIC50 melatonin and 1xIC50 cisplatin) was 6,7±0,17. Last, in GSH, mean expression in IC50 cisplatin group was 33,2±0,87, in IC50 melatonin group was 12,57±0,12, in IC50 doxorubicin group was 1,33±0,66, and in combination 1 group (1xIC50 melatonin and 1xIC50 cisplatin) was 11,73±0,67. There was significant difference of CTR-1 expression in combination 1 group which was higher (19.73%),  p-glycoprotein expression in combination 1 group which was lower (6,7%), and also GSH expression in combination 1 group was lower (11,73%) compared to other groups.
Conclusion: The group 1 combination of 1xIC50 melatonin and 1x IC50 cislatin with 1.841 mM and cisplatin 117.5 uM were able to reduce cisplatin chemotherapy resistance by increasing drug influx activity by increasing CTR-1 expression, decreasing drug efflux through decreasing p-glycoprotein expression, and decreased DNA repair activity through decreased GSH expression.