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Abstrak :

Kurkumin merupakan pigmen kuning alami dari rimpang kunyit yang diduga memiliki aktivitas kemopreventif terhadap sel kanker melalui mekanisme jalur pensinyalan apoptosis. Penelitian ini bertujuan untuk menguji hubungan kurkumin terhadap kadar protein RASSF1A,  Bax, dan aktivitas kaspase-3 dalam menunjang  mekanisme apoptosis pada sel kanker payudara CSA03, MCF-7, dan MDA-MB-468.

Penelitian eksperimen in vitro dilakukan di laboratorium terpadu Fakultas Kedokteran Universitas Indonesia Jakarta,  laboratorium terpadu Fakultas Kedokteran Universitas YARSI Jakarta, RSUPN Dr. Cipto Mangunkusumo Jakarta, serta RS Islam Jakarta tahun 2016–2018. Pemberian kurkumin terhadap sel kanker didasarkan atas perbedaan dosis dan waktu pemberian. Uji sitotoksisitas  setelah pemberian kurkumin ditentukan secara MTS.   Kadar protein RASSF1A dan Bax diuji secara ELISA. Aktivitas kaspase-3 digunakan untuk mengetahui apoptosis diuji secara flowsitometri. Selanjutnya perubahan morfologi sel diamati melalui pewarnaan acridine orange/ethidium bromide.

Pemberian kurkumin terhadap sel-sel yang diuji menunjukkan konsentrasi IC₅₀ yaitu 40,85 µg/mL pada sel CSA03; 75,73 µg/mL pada sel MCF-7; dan 380,79 µg/mL pada sel MDA-MB-468. Pemberian kurkumin menunjang mekanisme apoptosis melalui jalur RASSF1A, Bax, dan aktivitas kaspase-3 pada sel kanker payudara.

 

Kata Kunci:  Apoptosis, Bax, CSA03, kaspase-3, kurkumin,  MCF-7, MDA-MB-468, pewarnaan ganda, RASSF1A

 

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Curcumin is a natural yellow pigment from turmeric rhizome which is thought to have a chemopreventive effect on cancer through the mechanism of apoptotic signaling pathways. This study aims to examine the correlation of curcumin with protein level of RASSF1A, Bax, and caspase-3 activities in conjunction with the mechanism of apoptosis in CSA03, MCF-7, and MDA-MB-468 breast cancer cells.

In vitro experimental research was carried out at the Integrated Laboratory of Faculty of Medicine, Universitas Indonesia Jakarta; RSUPN. Dr. Cipto Mangunkusumo Jakarta; and Jakarta Islamic Hospital during 2016–2018. Curcumin was administered to the cancer cells in different doses and time. Cytotoxicity test after administration of curcumin was determined by MTS. The protein level of RASSF1A and Bax were measured by ELISA. Caspase-3 activity was used to determine apoptosis by flow cytometry. Furthermore, changes in cell morphology were observed by acridine orange/ethidium bromide staining.

The administration of curcumin to the cells showed IC50 concentrations of 40.85 µg/mL in CSA03 cells; 75.73 μg/mL in MCF-7 cells; and 380.79 µg/mL in MDA-MB-468 cells. The administration of curcumin supports the mechanism of apoptosis through the RASSF1A, Bax, and caspase-3 activity in breast cancer cells.

Abstrak :
[ABSTRAK
Pendahuluan: Di Indonesia belum ada penelitian tentang injeksi silikon dan komplikasinya, walaupun kasusnya banyak. Patogenesis granuloma silikon masih belum jelas. Beberapa penelitian mengemukakan peran sel T dan sitokin, namun belum ada yang meneliti tentang toleransi imun. Metode: Penelitian ini merupakan penelitian deskriptif analitik meliputi rancangan potong lintang membandingkan 3 kelompok, yaitu 31 jaringan granuloma dan 31 kulit submental pasien dengan suntikan silikon di dagu (kasus) dan 37 kulit normal (kontrol), terhadap gambaran klinis, histopatologis, dan respons imun melalui ekspresi sitokin TNF-a, IFN-g, IL-10, enzim IDO, serta sel Treg (CD4+CD25+); Penelitian eksperimental membiakkan darah penuh kasus dan orang normal, pada RPMI, dan RPMI yang distimulasi PHA, dan silikon. Dilanjutkan dengan mengukur kadar sitokin TNF-a, IFN-g, IL-10 dan IDO supernatan biakan darah. Penelitian dilakukan di klinik spesialis JMB, FMIPA, FKUI, FKUNAIR, dan lembaga Eijkman, tahun 2012 - 2014. Hasil Penelitian: Sebanyak 31 pasien granuloma akibat suntikan silikon di dagu umumnya datang berobat 12,5 tahun setelah penyuntikan, perubahan bentuk dagu terjadi pada tahun ke-4, perubahan warna pada tahun ke-5. Kadar sitokin proinflamasi di supernatan biakan darah lebih tinggi pada pasien granuloma daripada normal. Terdapat korelasi bermakna antara TNF-a di supernatan biakan darah dengan ekspresi TNF-a di jaringan granuloma. Enzim IDO, Treg, IL-10 di kulit submental berkorelasi bermakna dengan sitokin di granuloma. Sitokin anti inflamasi berperan pada kulit submental. Rasio TNF-a/IL-10 di supernatan biakan darah berkorelasi terbalik dengan ekspresi sel Treg di granuloma, membuktikan fungsi Treg sebagai toleransi imun, bekerja melalui IL-10. Enzim IDO di granuloma berkolerasi bermakna dengan rasio TNF-a/IL-10 di supernatan biakan darah dan Treg kulit submental. Simpulan: Enzim IDO bekerja sama dengan fungsi sel Treg dalam toleransi imun pada granuloma akibat suntikan silikon. TNF-a di supernatan biakan darah dan sitokin anti inflamasi di kulit submental dapat dijadikan prediktor untuk menilai respons imun yang terjadi akibat suntikan silikon.;

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ABSTRACT
Background: There is no study on silicone injections and its complications in Indonesia, yet, although the number of cases increased. The pathogenesis of silicone granulomas is still unclear. A few studies have been made to investigate the role of T cells and cytokines, however, none investigates the role of immune tolerance. Method: An analytical descriptive study encompassing cross sectional research was designed to compare 3 groups of 31 granuloma tissue and 31 submental skin of the patients with silicone injection in the chin (case) and 37 normal skin (control) on the clinical pictures, histopathological features and immune response through the expression of TNF-a, IFN-g, IL-10 cytokines, IDO enzyme, and Treg cells (CD4+CD25+). The experimental study cultured whole blood of the case and control patients and measured the level of TNF-a, IFN-g, IL-10 cytokines and IDO enzyme. The study was conducted in JMB specialist clinics, FMIPA, FKUI, FKUA, and Eijkman foundation from the year 2012 to 2014. Result: Thirty one patients with granuloma caused by silicone injection in the chin commonly seek medical advice 12.5 years after the injection, the chin shape changed on the fourth year and the skin color changed on the fifth year. Patients with granuloma had higher level of proinflammatory cytokines in their blood cultured supernatant. There was a significant correlation between TNF-a in blood cultured supernatant with the expression of TNF-a in the granuloma tissue. IDO enzyme, Treg cells, IL-10 in the submental skin significantly correlated with the cytokines in the granulomas. Anti inflammatory cytokines played a role on the submental skin. The ratio of TNF-a/IL-10 in blood cultured supernatant reversely correlated with the expression of Treg cells in the granuloma, demonstrating the function of Treg cells as an immune tolerance working through IL-10. IDO enzyme in the granulomas significantly correlated with the ratio of TNF-a/IL-10 in blood cultured supernatant and Treg in the submental skin. Conclusion: IDO enzyme collaborates with Treg cells in the immune tolerance caused by silicone injection. TNF-a in blood cultured supernatant and anti inflammatory cytokines in the submental skin can be utilized as predictors to assess the resulting immune response due to silicone injection. ;Background: There is no study on silicone injections and its complications in Indonesia, yet, although the number of cases increased. The pathogenesis of silicone granulomas is still unclear. A few studies have been made to investigate the role of T cells and cytokines, however, none investigates the role of immune tolerance. Method: An analytical descriptive study encompassing cross sectional research was designed to compare 3 groups of 31 granuloma tissue and 31 submental skin of the patients with silicone injection in the chin (case) and 37 normal skin (control) on the clinical pictures, histopathological features and immune response through the expression of TNF-a, IFN-g, IL-10 cytokines, IDO enzyme, and Treg cells (CD4+CD25+). The experimental study cultured whole blood of the case and control patients and measured the level of TNF-a, IFN-g, IL-10 cytokines and IDO enzyme. The study was conducted in JMB specialist clinics, FMIPA, FKUI, FKUA, and Eijkman foundation from the year 2012 to 2014. Result: Thirty one patients with granuloma caused by silicone injection in the chin commonly seek medical advice 12.5 years after the injection, the chin shape changed on the fourth year and the skin color changed on the fifth year. Patients with granuloma had higher level of proinflammatory cytokines in their blood cultured supernatant. There was a significant correlation between TNF-a in blood cultured supernatant with the expression of TNF-a in the granuloma tissue. IDO enzyme, Treg cells, IL-10 in the submental skin significantly correlated with the cytokines in the granulomas. Anti inflammatory cytokines played a role on the submental skin. The ratio of TNF-a/IL-10 in blood cultured supernatant reversely correlated with the expression of Treg cells in the granuloma, demonstrating the function of Treg cells as an immune tolerance working through IL-10. IDO enzyme in the granulomas significantly correlated with the ratio of TNF-a/IL-10 in blood cultured supernatant and Treg in the submental skin. Conclusion: IDO enzyme collaborates with Treg cells in the immune tolerance caused by silicone injection. TNF-a in blood cultured supernatant and anti inflammatory cytokines in the submental skin can be utilized as predictors to assess the resulting immune response due to silicone injection. , Background: There is no study on silicone injections and its complications in Indonesia, yet, although the number of cases increased. The pathogenesis of silicone granulomas is still unclear. A few studies have been made to investigate the role of T cells and cytokines, however, none investigates the role of immune tolerance. Method: An analytical descriptive study encompassing cross sectional research was designed to compare 3 groups of 31 granuloma tissue and 31 submental skin of the patients with silicone injection in the chin (case) and 37 normal skin (control) on the clinical pictures, histopathological features and immune response through the expression of TNF-a, IFN-g, IL-10 cytokines, IDO enzyme, and Treg cells (CD4+CD25+). The experimental study cultured whole blood of the case and control patients and measured the level of TNF-a, IFN-g, IL-10 cytokines and IDO enzyme. The study was conducted in JMB specialist clinics, FMIPA, FKUI, FKUA, and Eijkman foundation from the year 2012 to 2014. Result: Thirty one patients with granuloma caused by silicone injection in the chin commonly seek medical advice 12.5 years after the injection, the chin shape changed on the fourth year and the skin color changed on the fifth year. Patients with granuloma had higher level of proinflammatory cytokines in their blood cultured supernatant. There was a significant correlation between TNF-a in blood cultured supernatant with the expression of TNF-a in the granuloma tissue. IDO enzyme, Treg cells, IL-10 in the submental skin significantly correlated with the cytokines in the granulomas. Anti inflammatory cytokines played a role on the submental skin. The ratio of TNF-a/IL-10 in blood cultured supernatant reversely correlated with the expression of Treg cells in the granuloma, demonstrating the function of Treg cells as an immune tolerance working through IL-10. IDO enzyme in the granulomas significantly correlated with the ratio of TNF-a/IL-10 in blood cultured supernatant and Treg in the submental skin. Conclusion: IDO enzyme collaborates with Treg cells in the immune tolerance caused by silicone injection. TNF-a in blood cultured supernatant and anti inflammatory cytokines in the submental skin can be utilized as predictors to assess the resulting immune response due to silicone injection. ]