Pengaruh Triterpen Glikosida Teripang Timbu Kolong (Holothuria atra) terhadap Ekspresi Protein Fas dan Bcl-2 dalam Induksi Apoptosis pada Kultur Sel Supri’s Clone 1 (Sp-C1) sebagai Terapi Karsinoma Sel Skuamosa Lidah
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Institusion
Universitas Sumatera Utara
Author
Arma, Utmi
Subject
Apoptosis
Datestamp
2018-12-27 03:52:18
Abstract :
Squamous cell carcinoma of the oral cavity has the sixth highest incidence among various forms of cancer worldwide and eight of the most common cancers in the world. Squamous cell carcinoma of the oral cavity tends to increase and more than 90% of all oral cancers. Standard treatment is surgery, radiotherapy, chemotherapy or combination, but most patients with oral carcinoma, especially tongue carcinoma come in an advanced stage. Over the past few decades, there has been no effective systemic therapy for patients with advanced stage carcinoma, so the prognosis of patients with advanced carcinoma of the tongue is still poor. In carcinoesis, apoptotic dysregulation involving the Fas and Bcl-2protein expressiones is associated with tumor development, progression, and tumor resistance to therapy. One of the treatments being developed is the search for natural / herbal ingredients. This insight spurs the development of a therapeutic strategy targeted to activate apoptosis so that it can kill cancer cells. Triterpen glikosida contained in timbu cucumber has potency anticancer expected to induce apoptosis by intervening path involving protein expression of Fas and Bcl-2 The purpose of this study was to isolate the active compound triterpen glikosida of black sea cucumber and explore the effect of the active compound on the protein expression of Fas and Bcl-2 as the molecular mechanism of apoptosis to find molecular targets of tongue squamous cell carcinoma therapy.The study was a laboratory experimental study consisting of 2 stages. The first phase of the study was descriptive explorative to isolate the triterpen glycoside active compound from extract and fractionation of 3 types of sea cucumber (1)Black sea cucumber (Holothuria atra), (2)Bohadschia marmorata and (3) Stichopus variegatus cytotoxic test was performed as an ideal model of potential anticancer at that is a bioactive compound of 3 sea cucumbers that have the potential to kill cancer cells. The second phase of the study was an experimental in vitro study with complete randomized design (RAL) in the form of active triterpen glycoside trait test of trepang cucumber to assess its effect on Fas and Bcl-2 protein expression in induction of apoptosis on cell culture of Sp-C1. The study group consisted of a control group and a group of active triterpen glycoside compound trunks of cucumber dock with dose of 0.5xIC50, IC50, 1xIC50, 2xIC50 and 2.5xIC50 with observation time at 24 hours. Protein expression was measured by immunocytochemistry, while apoptotic detection Double Staining Ethidium Bromida Acridin Orange. The result of this research showed that active triterpen glycoside compound of trepang cucumber has a very strong anticancer activity (IC50 = 0,23 ?g / mL) to Sp-C1. The optimum protein expression of Fas and Bcl-2 for successful apoptosis initiation occurred at doses of 2xIC50 and 2.5xIC50 Keywords: apoptosis, protein expression of Fas and Bcl-2, Supri's-Clone 1 (Sp-C1), sea cucumber (Holothuria atra), triterpen glycoside, target therapy . There is a very strong relationship between expression of each of the Fas and Bcl-2 es with r values of 0.698 and 0.812 respectively with apoptosis r = 0.837. So the higher the dosage of active triterpenes of trusspole glikosida sea cucumber is higher then apoptosis in Sp-C1 culture. This indicates that the active triterpen glycoside compound of trepang cucumbers causes the e expression to produce a signal peak that activates the apoptotic phase of execution and shows the interaction between the extrinsic and intrinsic pathways thus strengthening the apoptotic induction process in inhibiting cancer growth. It was concluded that the active compound triterpenes of trunks of sea cucumber glycoside are very strong anticancer that can affect the protein expression of Fas and Bcl-2 e in inducing apoptosis.
Squamous cell carcinoma of the oral cavity has the sixth highest incidence among various forms of cancer worldwide and eight of the most common cancers in the world. Squamous cell carcinoma of the oral cavity tends to increase and more than 90% of all oral cancers. Standard treatment is surgery, radiotherapy, chemotherapy or combination, but most patients with oral carcinoma, especially tongue carcinoma come in an advanced stage. Over the past few decades, there has been no effective systemic therapy for patients with advanced stage carcinoma, so the prognosis of patients with advanced carcinoma of the tongue is still poor. In carcinoesis, apoptotic dysregulation involving the Fas and Bcl-2protein expressiones is associated with tumor development, progression, and tumor resistance to therapy. One of the treatments being developed is the search for natural / herbal ingredients. This insight spurs the development of a therapeutic strategy targeted to activate apoptosis so that it can kill cancer cells. Triterpen glikosida contained in timbu cucumber has potency anticancer expected to induce apoptosis by intervening path involving protein expression of Fas and Bcl-2 The purpose of this study was to isolate the active compound triterpen glikosida of black sea cucumber and explore the effect of the active compound on the protein expression of Fas and Bcl-2 as the molecular mechanism of apoptosis to find molecular targets of tongue squamous cell carcinoma therapy.The study was a laboratory experimental study consisting of 2 stages. The first phase of the study was descriptive explorative to isolate the triterpen glycoside active compound from extract and fractionation of 3 types of sea cucumber (1)Black sea cucumber (Holothuria atra), (2)Bohadschia marmorata and (3) Stichopus variegatus cytotoxic test was performed as an ideal model of potential anticancer at that is a bioactive compound of 3 sea cucumbers that have the potential to kill cancer cells. The second phase of the study was an experimental in vitro study with complete randomized design (RAL) in the form of active triterpen glycoside trait test of trepang cucumber to assess its effect on Fas and Bcl-2 protein expression in induction of apoptosis on cell culture of Sp-C1. The study group consisted of a control group and a group of active triterpen glycoside compound trunks of cucumber dock with dose of 0.5xIC50, IC50, 1xIC50, 2xIC50 and 2.5xIC50 with observation time at 24 hours. Protein expression was measured by immunocytochemistry, while apoptotic detection Double Staining Ethidium Bromida Acridin Orange. The result of this research showed that active triterpen glycoside compound of trepang cucumber has a very strong anticancer activity (IC50 = 0,23 ?g / mL) to Sp-C1. The optimum protein expression of Fas and Bcl-2 for successful apoptosis initiation occurred at doses of 2xIC50 and 2.5xIC50 Keywords: apoptosis, protein expression of Fas and Bcl-2, Supri's-Clone 1 (Sp-C1), sea cucumber (Holothuria atra), triterpen glycoside, target therapy . There is a very strong relationship between expression of each of the Fas and Bcl-2 es with r values of 0.698 and 0.812 respectively with apoptosis r = 0.837. So the higher the dosage of active triterpenes of trusspole glikosida sea cucumber is higher then apoptosis in Sp-C1 culture. This indicates that the active triterpen glycoside compound of trepang cucumbers causes the e expression to produce a signal peak that activates the apoptotic phase of execution and shows the interaction between the extrinsic and intrinsic pathways thus strengthening the apoptotic induction process in inhibiting cancer growth. It was concluded that the active compound triterpenes of trunks of sea cucumber glycoside are very strong anticancer that can affect the protein expression of Fas and Bcl-2 e in inducing apoptosis.
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