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Ekspresi Relatif mRNA Hypoxia Inducible Factor-1? pada Sel Glioma Penderita Hardiany, Novi Silvia; Sadikin, Mohamad; Wanandi, Septelia Inawati
Indonesian Journal of Cancer Vol 7, No 1 (2013): Jan - Mar 2013
Publisher : "Dharmais" Cancer Center Hospital

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (530.441 KB)

Glioma merupakan tumor otak primer yang cukup sering ditemukan di Indonesia di antara keganasan otak lainnya. Penderita glioma mempunyai angka harapan hidup yang rendah dikarenakan seringnya mengalami resistansi terapi. Hipoksia merupakan salah satu faktor yang mempengaruhi respons terapi radiasi. Penelitian ini menganalisis ekpresi mRNA HIF-1?, sebagai petanda jaringan hipoksia, pada glioma derajat rendah dibandingkan dengan glioma derajat tinggi. Sampel terdiri dari 15 jaringan glioma derajat rendah, 6 jaringan glioma derajat tinggi, dan 21 sel lekosit dari penderita glioma sebagai kontrol sel normal. Ekspresi relatif mRNA HIF-1? ditentukan secara kuantitatif dengan menggunakan Real Time RT-PCR. Hasil penelitian menunjukkan bahwa ekspresi mRNA HIF-1? pada glioma derajat tinggi lebih tinggi dibandingkan dengan glioma derajat rendah, namun tidak bermakna secara statistik. Hal yang menarik yaitu ditemukan peningkatan ekspresi mRNA HIF-1? dengan nilai yang bervariasi pada 73 % sampel glioma derajat rendah dibandingkan dengan sel lekosit sebagai kontrol sel normal. Begitu juga pada glioma derajat tinggi ditemukan peningkatan mRNA HIF-1? dengan nilai yang bervariasi pada 100% sampel. Dengan demikian, dapat disimpulkan bahwa terdapat variasi derajat hipoksia pada glioma derajat rendah dan derajat tinggi yang mungkin mempengaruhi keberhasilan terapi radiasi.Kata kunci: Glioma, mRNA HIF-1?, hipoksia

Kurkumin Meningkatkan Sensitivitas Sel Kanker Payudara terhadap Tamoksifen melalui Penghambatan Ekspresi P-glikoprotein dan Breast Cancer Resistance Protein Sianipar, Erlia Anggrainy; Louisa, Melva; Wanandi, Septelia Inawati
Jurnal Farmasi Indonesia Vol 9, No 2 (2017)
Publisher : Indonesian Research Gateway

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35617/jfi.v9i2.578

Background:The decreased of sensitivity or resistance to tamoxifen occured after long-term treatment in breast cancer. One of the major factor in tamoxifen resistance is over expression of efflux transporter P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP). Curcumin has known as inhibitor of P-gp and BCRP. The addition of curcumin to the tamoxifen resistant cells is expected to increase the sensitivity of breast cancer cells to tamoxifen. Method:MCF-7 breast cancer cell linewas induced with tamoxifen 1 uM for 10 passage (MCF-7(T)), then cell viability and mRNA expression of P-gp and BCRP were analyzed. To the MCF- 7(T) cells, curcumin was given at of 5/10/20 uM with or without tamoxifen for 5 days and cell viability and mRNA expression of P-gp and BCRP were analyzed on day 5. As positive control, verapamil 50 uM was used as P-gp inhibitor, ritonavir 15 yM and nelfinavir 15 yM were used as BCRP inhibitor. Result:MCF-7(T) cells sensitivity to tamoxifen has decreased with 11,8 times reduction in sensitivity towards tamoxifen, the increased of cell viability, and mRNA expression of P-gp and BCRP mRNA increased 10,82 and 4,04 fold respectively in MCF-7(T) cells. Then after administration of curcumin with or without tamoxifenfor5 days then the cell viability and the mRNA expression of P-gp mRNA and BCRP decreased. Conclusion:Curcumin iIncreased the sensitivity of MCF-7(T) to tamoxifen, characterized by the decreased of cell viability and mRNA expression of P-gp and BCRP. But the administration of combination curcumin with tamoxifen was more potent than curcumin alone. The iIncreased sensitivity was estimated at least in part through the inhibition of P-gp and BCRP MRNA expression by curcumin. Background:The decreased of sensitivity or resistance to tamoxifen occured after long-term treatment in breast cancer. One of the major factor in tamoxifen resistance is over expression of efflux transporter P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP). Curcumin has known as inhibitor of P-gp and BCRP. The addition of curcumin to the tamoxifen resistant cells is expected to increase the sensitivity of breast cancer cells to tamoxifen. Method:MCF-7 breast cancer cell linewas induced with tamoxifen 1 uM for 10 passage (MCF-7(T)), then cell viability and mRNA expression of P-gp and BCRP were analyzed. To the MCF- 7(T) cells, curcumin was given at of 5/10/20 uM with or without tamoxifen for 5 days and cell viability and mRNA expression of P-gp and BCRP were analyzed on day 5. As positive control, verapamil 50 uM was used as P-gp inhibitor, ritonavir 15 yM and nelfinavir 15 yM were used as BCRP inhibitor. Result:MCF-7(T) cells sensitivity to tamoxifen has decreased with 11,8 times reduction in sensitivity towards tamoxifen, the increased of cell viability, and mRNA expression of P-gp and BCRP mRNA increased 10,82 and 4,04 fold respectively in MCF-7(T) cells. Then after administration of curcumin with or without tamoxifenfor5 days then the cell viability and the mRNA expression of P-gp mRNA and BCRP decreased. Conclusion:Curcumin iIncreased the sensitivity of MCF-7(T) to tamoxifen, characterized by the decreased of cell viability and mRNA expression of P-gp and BCRP. But the administration of combination curcumin with tamoxifen was more potent than curcumin alone. The iIncreased sensitivity was estimated at least in part through the inhibition of P-gp and BCRP MRNA expression by curcumin.

HDAC2 and PCNA expression is correlated to decreasing of endoxifen sensitivity in human breast cancer stem cells ALDH+ Syarifah Dewi; Mohamad Sadikin; Muchlis Ramli; Septelia Inawati Wanandi
Health Science Journal of Indonesia Vol 10 No 2 (2019)
Publisher : Sekretariat Badan Penelitian dan Pengembangan Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22435/hsji.v12i2.2449

Latar belakang: Sel punca kanker payudara (breast cancer stem cells/BCSC) adalah subpopulasi sel kanker yang memiliki kemampuan menghasilkan tumor baru dan bersifat seperti sel punca. Penelitian kami sebelumnya menggunakan jaringan kanker payudara mengungkapkan bahwa ekspresi gen histone deacetylase 2 (HDAC2) dan proliferating cell nuclear antigen (PCNA) ditemukan perbedaan signifikan setelah terapi neoajuvan hormon dan kemoterapi. Penelitian ini bertujuan untuk menganalisis hubungan antara ekspresi HDAC2 dan PCNA dengan kelangsungan hidup sel punca kanker payudara dengan penanda aldehyde dehydrogenase + (ALDH+) yang diberi perlakuan endoksifen. Metode: Sampel adalah BCSC primer manusia ALDH+ yang diberi perlakuan endoksifen 4 uM masingmasing selama 2, 4, 6, 8, 10, 12, 14 hari. Viabilitas sel dilihat dengan menggunakan trypan blue dan ekspresi mRNA HDAC2 dan PCNA ditentukan menggunakan qRT-PCR. Hasil: Viabilitas BCSCs ALDH + menurun setelah 2 sampai 4 hari pemberian endoksifen. Pada periode ini juga didapatkan ekspresi mRNA HDAC2 dan PCNA mengalami penurunan. Tetapi setelah pemberian endoksifen selama 8 hari, viabilitas BCSCs ALDH + mengalami peningkatan dan ditemukan peningkatan yang signifikan pada hari ke-14 pemberian endoksifen. Ekspresi mRNA HDAC2 dan PCNA juga menunjukkan peningkatan mulai pada hari ke-8 dan terus meningkat hingga hari ke-14 pemberian endoksifen. Penelitian ini menunjukkan pola yang sama antara ekspresi mRNA HDAC2 dan PCNA dan viabilitas sel. Kesimpulan: Induksi endoksifen yang lama menurunkan sensitivitas efek endoksifen pada BCSC manusia dan ekspresi HDAC2 dan PCNA berkorelasi dengan viabilitas BCSC manusia setelah induksi endoksifen. (Health Science Journal of Indonesia 2019;10(2):77-81) Kata kunci: sel punca kanker payudara, viabilitas sel, HDAC2, PCNA, endoksifen Abstract Background: Breast cancer stem cells (BCSCs) are subpopulation of cancer cells that has the ability to generate new tumor and similar properties to stem cell. Our previous study using breast cancer patients revealed that gene expression of histone deacetylase 2 (HDAC2) and proliferating cell nuclear antigen (PCNA) were significantly altered after neoadjuvant hormone and chemotherapy. This study aimed to analyze the correlation between HDAC2 and PCNA expressions with the viability of breast cancer stem cells aldehyde dehydrogenase + (BCSC ALDH+) treated by endoxifen. Method: Samples are human primary BCSCs ALDH+ that treated with 4 uM of endoxifen for 2, 4, 6, 8, 10, 12, 14 days, respectively. Cell viability was measured using trypan blue exclusion assay and the mRNA expressions of HDAC2 and PCNA were determined using qRT-PCR. Results: The viability of BCSCs ALDH+ was decreased after 2 days until 4 days-endoxifen treatment. It also demonstrated that mRNA expression of HDAC2 and PCNA were decreased in this period. But after 8 days endoxifen treatment, the viability of BCSCs ALDH+ was increased. The increasing of viability was higher in 14 days-endoxifen treatment. The mRNA expression of HDAC2 and PCNA also showed increasing begin on 8 days and continued to increase until 14-days endoxifen treatment. We found a similar pattern between HDAC2 and PCNA expression and cell viability. Conclusion: Prolonge endoxifen treatment decrease sensitivity of endoxifen effect in human BCSC and the expression of HDAC2 and PCNA are correlated to human BCSCs viability after endoxifen treatment. (Health Science Journal of Indonesia 2019;10(2):77-81) Keywords: human breast cancer stem cells, viability, HDAC2, PCNA, endoxifen

Ekspresi Relatif mRNA Hypoxia Inducible Factor-1? pada Sel Glioma Penderita Novi Silvia Hardiany; Mohamad Sadikin; Septelia Inawati Wanandi
Indonesian Journal of Cancer Vol 7, No 1 (2013): Jan - Mar 2013
Publisher : National Cancer Center - Dharmais Cancer Hospital

Glioma merupakan tumor otak primer yang cukup sering ditemukan di Indonesia di antara keganasan otak lainnya. Penderita glioma mempunyai angka harapan hidup yang rendah dikarenakan seringnya mengalami resistansi terapi. Hipoksia merupakan salah satu faktor yang mempengaruhi respons terapi radiasi. Penelitian ini menganalisis ekpresi mRNA HIF-1?, sebagai petanda jaringan hipoksia, pada glioma derajat rendah dibandingkan dengan glioma derajat tinggi. Sampel terdiri dari 15 jaringan glioma derajat rendah, 6 jaringan glioma derajat tinggi, dan 21 sel lekosit dari penderita glioma sebagai kontrol sel normal. Ekspresi relatif mRNA HIF-1? ditentukan secara kuantitatif dengan menggunakan Real Time RT-PCR. Hasil penelitian menunjukkan bahwa ekspresi mRNA HIF-1? pada glioma derajat tinggi lebih tinggi dibandingkan dengan glioma derajat rendah, namun tidak bermakna secara statistik. Hal yang menarik yaitu ditemukan peningkatan ekspresi mRNA HIF-1? dengan nilai yang bervariasi pada 73 % sampel glioma derajat rendah dibandingkan dengan sel lekosit sebagai kontrol sel normal. Begitu juga pada glioma derajat tinggi ditemukan peningkatan mRNA HIF-1? dengan nilai yang bervariasi pada 100% sampel. Dengan demikian, dapat disimpulkan bahwa terdapat variasi derajat hipoksia pada glioma derajat rendah dan derajat tinggi yang mungkin mempengaruhi keberhasilan terapi radiasi.Kata kunci: Glioma, mRNA HIF-1?, hipoksia

Characterization of Thermoplasma Species Cultured from Sampling on Tangkuban Perahu, Indonesia AMARILA MALIK; IMAN SANTOSO; ANDI YEHUDA; SERUNI K.U. FREISLEBEN; SEPTELIA INAWATI WANANDI; HARALD HUBER; ZESSINDA LUTHFA; ROSARI SALEH; HANS-JOACHIM FREISLEBEN
Microbiology Indonesia Vol. 8 No. 1 (2014): March 2014
Publisher : Indonesian Society for microbiology

Archaea is an organisme with unique feature because of its ability to inhabit an extremophyle conditions. Our expeditions to Tangkuban Perahu, West Java aimed to obtain archaealstrains from the solfatara fields located in Domas crater. From the samples, we intended to culture Thermoplasma species growing around 55 Â°C below pH 2, which until now have not yet been fully characterized. We collected five samples from mud holes with temperatures from 52 Â°C to 57 Â°C and pH below 2. In serial cultures of up to 8 transfers in Freundtâ€™s medium we grew tetraetherlipid synthesizing Thermoplasma species as documented by phase contrast microscope. Total membrane lipid extracts were analysed by thin layer chromatography; the pattern matched total lipid extracts from Thermoplasma acidophilum DSM 1728 membranes. For confirmation, 16S rDNA identification performing PCR and sequencing were carried-out. Analysis using BLAST showed T. acidophilum identities as the highest similarity of 99%, followed by T. volcanium, also with99% similarity (ANKF776908 and ANKF776909). This is the first report of culturing cell-wall-less thermoacidophilicarchaea,in particular Thermoplasma species in Indonesian laboratories.

EKSPRESI RELATIF mRNA HIF-1α PADA JANTUNG, OTAK DAN DARAH TIKUS SELAMA INDUKSI HIPOKSIA SISTEMIK Wanandi, Septelia Inawati; Dewi, Syarifah; Paramita, Reni
Makara Journal of Science Vol. 13, No. 2
Publisher : UI Scholars Hub

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Relative Expression of HIF-1α mRNA in Rat Heart, Brain and Blood During Induced Systemic Hypoxia. Hypoxia is a pathological condition in which the body as a whole or region of the body (tissue or cell) deprived of adequate oxygen supply. The transcriptional regulator hypoxia inducible factor-1 (HIF-1) is an essential mediator of O2 homeostasis. Unlike the β sub unit (HIF-1β), the activity of HIF-1α is controlled in an oxygen-dependent manner. It has been reported that the stability and expression of HIF-1α during hypoxia is remarkably higher than those under normoxic conditions.The aim of this study was to analyze the adaptive tissue responses during induced systemic hypoxia by comparation of relative expression of mRNA HIF-1α in rat heart, brain and blood. Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia by placing them in the hypoxic chamber supplied by 8-10% of O2 for 0, 1, 7, 14 and 21 days, respectively. The relative expression level of HIF-1α mRNA in brain, heart and leucocyte cells were analyzed using quantitative RT-PCR assay (Real Time PCR) based on Pfaff’s formula. This study demonstrates that the increased of relative expression of HIF-1α mRNA during induced systemic hypoxia reached its maximum level at day 7 (in heart) or at day 14 (in brain), whereas in leucocyte cells the stimulation of HIF-1α expression was intensively maintained up to 21 days although the expression has reached the remarkably high level. We could conclude that HIF-1α as an oxygen sensing during systemic hypoxia has different capacity and sensitivity in brain, heart and blood tissues, due to the importance of oxygen homeostasis in each tissue

EKSPRESI RELATIF mRNA HIF-1α PADA JANTUNG, OTAK DAN DARAH TIKUS SELAMA INDUKSI HIPOKSIA SISTEMIK Wanandi, Septelia Inawati; Dewi, Syarifah; Paramita, Reni
Makara Journal of Science Vol. 13, No. 2
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Relative Expression of HIF-1α mRNA in Rat Heart, Brain and Blood During Induced Systemic Hypoxia. Hypoxia is a pathological condition in which the body as a whole or region of the body (tissue or cell) deprived of adequate oxygen supply. The transcriptional regulator hypoxia inducible factor-1 (HIF-1) is an essential mediator of O2 homeostasis. Unlike the β sub unit (HIF-1β), the activity of HIF-1α is controlled in an oxygen-dependent manner. It has been reported that the stability and expression of HIF-1α during hypoxia is remarkably higher than those under normoxic conditions.The aim of this study was to analyze the adaptive tissue responses during induced systemic hypoxia by comparation of relative expression of mRNA HIF-1α in rat heart, brain and blood. Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia by placing them in the hypoxic chamber supplied by 8-10% of O2 for 0, 1, 7, 14 and 21 days, respectively. The relative expression level of HIF-1α mRNA in brain, heart and leucocyte cells were analyzed using quantitative RT-PCR assay (Real Time PCR) based on Pfaff’s formula. This study demonstrates that the increased of relative expression of HIF-1α mRNA during induced systemic hypoxia reached its maximum level at day 7 (in heart) or at day 14 (in brain), whereas in leucocyte cells the stimulation of HIF-1α expression was intensively maintained up to 21 days although the expression has reached the remarkably high level. We could conclude that HIF-1α as an oxygen sensing during systemic hypoxia has different capacity and sensitivity in brain, heart and blood tissues, due to the importance of oxygen homeostasis in each tissue

Andrographis paniculata Ethanolic Extract Improved Doxorubicin-induced Cardiac Inflammation, Alterations in Liver Function Parameters and Anemia Eziefule, Oluebube Magnificient; Arozal, Wawaimuli; Wanandi, Septelia Inawati; Louisa, Melva; Wuyung, Puspita Eka; Dewi, Syarifah; Nafrialdi, Nafrialdi; Dewi, Yulia Ratna; Nabillah, Deya Adiby
Molecular and Cellular Biomedical Sciences Vol 8, No 2 (2024)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v8i2.444

Background: Doxorubicin (DOX), an efficacious chemotherapy drug is compromised by cardiotoxicity, myelosuppression, and hepatotoxicity. Due to the limited success of current treatments for DOX toxicity, there is a pressing need to explore alternative medical interventions, particularly from plant sources. This study was conducted to investigate the potential protective effect of ethanolic extract of Andrographis paniculata leaves (EEAP) against DOX-induced cardiac inflammation, liver toxicity, and anemia.Materials and methods: Sprague-Dawley rats were intraperitoneally injected with DOX at a total dose of 16 mg/kgBW. EEAP was administered orally for 4 weeks at doses of 125, 250, and 500 mg/kgBW/day according to the assigned treatment groups. The mRNA expression levels of interleukin-1β (IL-1β) and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) in the heart tissue, along with the concentrations of nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) and calcium level were examined. Additionally, the hematological parameters (including hematocrit, hemoglobin and red blood cells (RBCs)), aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), and malondialdehyde (MDA) levels in blood were also analyzed.Results: EEAP dose-dependently decreased the mRNA expressions of IL-1β (p<0.05), tended to decrease mRNA expression of NLRP3 and the concentrations of NFκB and calcium in heart tissue compared with the DOX-only group. Additionally, EEAP dose-dependently decreased ALP values (p<0.0001) and tended to improve hematological parameters, as well as AST and MDA levels in serum.Conclusion: This extract may prevent DOX-induced cardiac inflammation, anemia, and hepatotoxicity. However, further studies are needed to confirm these findings, including the efficacy profile of the extract in cancer rats treated with DOX.Keywords: doxorubicin, Andrographis paniculata, inflammation, anemia, hepatotoxicity, herbal medicine

Evasion of the Immune System by Glioblastoma Multiforme: An Obstacle to Achieving Effective Therapies Kawengian, Kevin Johanes; Wanandi, Septelia Inawati
Molecular and Cellular Biomedical Sciences Vol 8, No 2 (2024)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v8i2.434

Glioblastoma multiforme (GBM), a highly aggressive and malignant form of brain cancer, continues to pose a significant challenge in the field of oncology. Despite ongoing advancements in treatment strategies, the prognosis for GBM patients remains grim, with a 5-year survival rate hovering around 5%. The management of GBM involves multiple therapeutic approaches, including immunotherapy, but optimal treatment outcomes in terms of overcoming tumor recurrence and resistance have not been achieved. A key factor contributing to therapy resistance and the progression of GBM is the tumor's ability to evade the immune system, referred to as immune escape from cancer. This phenomenon reflects the tumor cells' efforts to adapt and survive the body's immune response. The release and expression of molecules like TGF-ß, IL-10, PD-L1, and NKG2DL by GBM cells impact the activation, recognition, and elimination of tumor cells by the immune system. Additionally, the involvement of cells such as MDSCs, Tregs, and TAMs plays a role in inhibiting the immune system's function, thereby promoting the development of GBM cells. A better comprehension of GBM's immune escape, supported by technological advances, will significantly aid in the future management of GBM patients' treatment.Keywords: glioblastoma multiforme, GBM, cancer immunity, immune evasion, immune escape, immunotherapy

MnSOD Gene Knockout Promotes Apoptosis in Triple-Negative Breast Cancer BT-549 Cells Through Survivin Inhibition and Caspase-3 and Caspase-9 Modulation Ghanny, Niken Rahmah; Wanandi, Septelia Inawati; Arumsari, Sekar; Paramita, Rafika Indah; Syahrani, Resda Akhra; Putri, Putu Indah Paramita Adi
Makara Journal of Science
Publisher : UI Scholars Hub

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Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype characterized by the lack of estro-gen, progesterone, and human epidermal growth factor receptor 2 (HER2) receptors. TNBC cells are becoming more aggressive due to the high expression of manganese superoxide dismutase (MnSOD) antioxidants to suppress reactive oxygen species-induced apoptosis and promote oncogenic signaling. This study was aimed to evaluate the effects of MnSOD knockout (KO) on TNBC cell apoptosis by assessing the survivin, caspase-9, and caspase-3 expressions. BT-549 TNBC cells containing the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-edited MnSOD gene were used to evaluate the messenger RNA (mRNA) expressions of survivin using the RT-qPCR assay. The West-ern blot assay was used to measure protein expressions of survivin, caspase-9, and caspase-3. Interactions between MnSOD and apoptosis-related proteins were simulated using computational methods based on molecular docking analysis and protein−protein interaction network. This study revealed that MnSOD KO decreased the binding affinity between MnSOD and survivin, in line with the significant reduction of survivin mRNA and protein expressions whereas the protein expressions of caspase-9 and caspase-3 increased in MnSOD KO cells. Therefore, MnSOD plays a pivotal role in BT-549 cell apoptosis by modulating survivin, caspase-9, and caspase-3 gene expressions. This study provides insights into a novel therapeutic strategy to mitigate the aggressiveness of TNBC by disrupting MnSOD gene expression. Further studies should elaborate the MnSOD signaling pathways involving closely related apoptotic proteins.

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