Kajian In Vitro dan In Silico Daun Sirih Merah sebagai Antikanker Kolon pada Sel WiDr serta Proliferator Sel Limfosit.

Abstract

Kanker kolon atau Kanker kolorektal adalah suatu tumor maligna yang muncul dari jaringan epitel dari kolon dan/atau rektum. Secara global, kanker kolon masih merupakan salah satu masalah kesehatan utama. Salah satu tanaman yang berpotensi mengurangi keganasan kanker kolon adalah sirih merah (Piper crocatum Ruiz & Pav). Fraksi etil asetat dari ekstrak etanol sirih merah diketahui memiliki aktivitas antioksidan tertinggi pada beberapa uji antioksidan. Penelitian ini bertujuan menguji secara in vitro fraksi etil asetat ekstrak etanol daun sirih merah sebagai antikanker kolon pada sel WiDr dan proliferator limfosit. Selain itu, penelitian ini juga bertujuan untuk mengetahui mekanisme antikanker kolon secara in silico pada reseptor human leukotriene A4 hydrolase, BCL 2 navitoclax analog dan chek1. Penelitian in vitro dengan menggunakan uji MTT untuk antikanker dan proliferasi limfosit. Penelitian in silico menggunakan software YASARA Structure dengan 3 reseptor antikanker kolon, yaitu: 4LXD, 3U9W dan 2R0U. Hasil penelitian in vitro menunjukkan fraksi etil asetat ekstrak etanol daun sirih merah dapat menghambat pertumbuhan sel kanker kolon WiDr dengan rata rata tertinggi 98,18 ± 1,21 % pada konsentrasi 500 ppm dan meningkatkan proliferasi limfosit dengan rata-rata tertinggi 143,3 ± 19,46 % pada konsentrasi 20 ppm. Hasil penelitian in silico menunjukkan ligan 4-(4-methoxy-phenylamino)-2,3- dihydro-1H-4a,9-diaza-cyclopenta (b) fluorine-10- carbonitrile memiliki interaksi terbaik dengan BCL 2 navitoclax analog dengan energi bebas ikat sebesar -8,4470 Kcal/mol, nilai konstanta inhibisi (Ki) sebesar 6,33 x 10-7 μM dengan 7 residu asam amino (Tyr105, Met112, Glu133, Leu134, Gly142, Arg143, dan Ala146) yang berinteraksi pada sisi aktif reseptor. Reseptor human leukotriene A4 hydrolase berinteraksi terbaik dengan ligan N-1,N-9-Bis [E-(2- nitrophenyl)methylene] non anedihydrazide (CID: 9595915) dengan energi bebas ikat sebesar -11,3160 Kcal/mol, nilai Ki sebesar 4,97 × 10-9 μM dan berinteraksi terhadap 14 residu (Gln1136, Tyr1267, Gly1269, Met1270, His1295, Glu1296, Trp131, Phe1314, Val1367, Pro1374, Ala1377, Tyr1378, Pro1382, Tyr1383) terhadap sisi aktif. Reseptor chek1 memiliki interaksi terbaik dengan senyawa 4-({4,6-Bis[3R,5S)-3,5-diamino-1-piperydinyl] -1,3,5-triazin-2-yl} amino benzenosulfonamide dengan energi bebas ikat sebesar -9,7110 Kcal/mol, dengan nilai Ki 7,48 × 10-8 μM dan interaksi terhadap 10 residu asam amino (Leu15, Val23, Lys38, Leu84, Tyr86, Cys87, Gly90, Glu91, Glu134, dan Asn135) pada sisi aktif. Hal ini menunujukkan bahwa senyawa bioaktif fraksi etil asetat ekstrak etanol daun sirih merah memiliki aktivitas antikanker kolon terhadap sel WiDr dan bertindak sebagai proliferator sel limfosit.

Colon cancer or colorectal cancer is a malignant tumor that arises from the epithelial tissue of the colon and/or rectum. Globally, colon cancer is still one of the major health problems. One plant that has the potential to reduce colon cancer malignancy is red betel (Piper crocatum Ruiz & Pav). The ethyl acetate fraction of red betel ethanol extract is known to have the highest antioxidant activity in several antioxidant tests. This study aimed to examine in vitro of the ethyl acetate fraction of red betel leaf ethanol extract as a colon anticancer on WiDr cells and as lymphocyte proliferator. In addition, this study also aimed to determine the mechanism of colon anticancer in silico againts human leukotriene A4 hydrolase, BCL- 2 navitoclax analog, and chek1 receptor. In vitro research using the MTT assay for anticancer and lymphocyte proliferation analysis. In silico research using YASARA Structure software with 3 colon anticancer receptors, namely: 4LXD, 3U9W and 2R0U. The results of in vitro research showed that the ethyl acetate fraction of red betel leaf ethanol extract could inhibit the growth of WiDr colon cancer cells with the highest average of 98.18 ± 1.21% at a concentration of 500 ppm and could increase lymphocyte proliferation with the highest average of 143.3 ± 19 ,46 % at a concentration of 20 ppm. The results of the in silico study showed that the ligand 4-(4-methoxy-phenylamino)-2,3-dihydro-1H-4a,9-diaza-cyclopenta (b) fluorine 10-carbonitrile had the best interaction with BCL 2 navitoclax analog with binding free energy of -8.4470 Kcal/mol, constant inhibition value (Ki) of 6.33 x 10-7 μM, interacted with 7 amino acid residues (Tyr105, Met112, Glu133, Leu134, Gly142, Arg143, and Ala146) on the receptor's active site. Human leukotriene A4 hydrolase receptor interacted best with the ligand N 1,N-9-Bis [E-(2-nitrophenyl)methylene] non-anedihydrazide (CID: 9595915) with binding free energy of -11,3160 Kcal/mol, Ki value of 4.97 × 10-9 μM and interacted with 14 residues (Gln1136, Tyr1267, Gly1269, Met1270, His1295, Glu1296, Trp131, Phe1314, Val1367, Pro1374, Ala1377, Tyr1378, Pro1382, Tyr1383) on the active site . The chek1 receptor had the best interaction with the compound 4-({4,6-Bis[3R,5S)-3,5-diamino-1-piperydinyl]-1,3,5-triazine-2-yl} am ino benzenosulfonamide with a binding free energy of -9,7110 Kcal/mol, with a Ki value of 7.48 × 10-8 μM, interacted with 10 amino acid residues (Leu15, Val23, Lys38, Leu84, Tyr86, Cys87, Gly90, Glu91, Glu134, and Asn135) on the active side. This shows that the bioactive compounds conceived by the ethyl acetate fraction of red betel leaf ethanol extract have colon anticancer activity againts WiDr cells and act as lymphocyte cell proliferator.