Tertiana, Nur Iedha and dr. Happy Kurnia Permatasari, Ph.D and dr. Hikmawan Wahyu Sulistomo, Ph.D (2023) Pengaruh Caulerpa racemosa Terhadap Ekspresi Bcl-2, BAX, Apoptosis dan Viabilitas pada Kultur Sel Kanker HeLa. Magister thesis, Universitas Brawijaya.
Abstract
Pendahuluan: Pada wanita, kanker serviks menduduki peringkat kedua kanker yang sering terjadi setelah kanker payudara. Penyebab utama kanker serviks adalah infeksi tanpa gejala, persisten, atau kronis oleh satu atau lebih Human Papilloma Virus (HPV) onkogenik. Pada tingkat molekuler, terjadi integrasi genom virus ke dalam genom inang yang menyebabkan overekspresi protein E6 dan E7 dari HPV. Peningkatan tersebut menyebabkan diferensiasi dan replikasi virus serta degradasi p53. Gen supresor tumor p53 mengatur ekspresi Bcl-2 dan BAX. Upregulasi protein antiapoptosis Bcl-2 dan hilangnya BAX adalah metode penghindaran yang dominan. Caulerpa memiliki metabolit bioaktif yaitu alkaloid, terpenoid, flavonoid, steroid dan tanin dan telah dilaporkan bioaktivitasnya terhadap banyak penyakit termasuk kanker. Belum ada penelitian mengenai manfaat Caulerpa racemosa yang berasal dari perairan Indonesia. Selain itu belum jelas mekanisme pensinyalan apoptosis yang menjadi target dari Caulerpa racemosa. Metode: Eksperimental murni menggunakan rancangan randomized post test controlled group design. Sampel yang digunakan adalah kultur sel HeLa. Jumlah perlakuan dibagi menjadi 4 kelompok, yakni kontrol (tanpa terapi), kelompok 1 (terapi C. racemosa 50 μg/mL), kelompok 2 (terapi C. racemosa 100 μg/mL), kelompok 3 (terapi C. racemosa 200 μg/mL). Viabilitas sel menggunakan trypan blue exclusion assay. Apoptosis menggunakan flowcytometry. Ekspresi Bcl-2 dan BAX terlihat pada pengecatan imuunofluoresence. Hasil: Pada viabilitas 24 jam, didapatkan p=0.018 (Kruskal-Wallis). Uji spearman mengindikasikan hubungan negatif sempurna (p=0,002, r= -0,802). Pada viabilitas 48 jam p=0,015 (Kruskal-Wallis). Uji Spearman mengindikasikan hubungan negatif sempurna (p=0,000, r= -0,975). Dilakukan uji Dunn’s Post Hoc sebagai uji lanjutan. Apoptosis diperoleh hasil p=0,023 (Kruskal-Wallis). Uji Spearman menunjukkan korelasi positif sempurna (p=0,000, r=0,908). Uji ANOVA terhadap BAX diperoleh perbedaan bermakna antar kelompok (p=0.000). Uji Tukey HSD menunjukkan terdapat perbedaan signifikan (p<0,05). Uji Pearson menunjukkan hubungan positif kuat antara dosis dengan intensitas BAX (p=0,000, r-0.880). Uji ANOVA terhadap Bcl-2 diperoleh perbedaan bermakna antar kelompok (p=0.000). Uji Tukey HSD menunjukkan perbedaan nilai signfkansi pada kelompok perlakuan (p<0,05). Uji Pearson megindikasikan secara statistik, tidak terdapat hubungan yang bermakna antara dosis dan intensitas Bcl-2 dengan korelasi positif lemah (p=0,435, r=0,249). Kesimpulan: Ekstrak C. racemosa dapat menurunkan viabilitas, menginduksi apoptosis, meningkatan ekspresi BAX. Peningkatan ekspresi dari Bcl-2 kultur sel merupakan mekanisme pertahanan sel dan diimbangi dengan peningkatan ekspresi BAX sehingga sel tetap memasuki fase apoptosis.
English Abstract
Introduction: In women, cervical cancer is the second most common cancer after breast cancer. The main cause of cervical cancer is asymptomatic, persistent, or chronic infection by one or more oncogenic Human Papilloma Virus (HPV). At the molecular level, integration of the viral genome into the host genome results in overexpression of the E6 and E7 proteins of HPV. This increase causes differentiation and viral replication and degradation of p53. The p53 tumor suppressor gene regulates Bcl-2 and BAX expression. Upregulation of the antiapoptotic protein Bcl-2 and loss of BAX are the predominant avoidance methods. Caulerpa has bioactive metabolites namely alkaloids, terpenoids, flavonoids, steroids and tannins and has been reported for its bioactivity against many diseases including cancer. There has been no research on the benefits of Caulerpa racemosa originating from Indonesian waters. In addition, it is unclear which mechanism of apoptotic signaling is the target of Caulerpa racemosa. Methods: True experimental study using a randomized post test controlled group design. The sample used was HeLa cell culture. The number of treatments was divided into 4 groups, control (without therapy), group 1 (therapy with C. racemosa 50 μg/mL), group 2 (therapy with C. racemosa 100 μg/mL), group 3 (therapy with C. racemosa 200 μg/mL). mL). Observation for cell viability using trypan blue exclusion assay. Apoptosis using flowcytometry. The expression of Bcl-2 and BAX was seen in the immunofluorescence staining. Results: At 24 hours of viability, p=0.018 (Kruskal-Wallis). Spearman's test showed a perfect negative relationship (p=0.002, r= -0.802). At 48 hours viability p=0.015 (Kruskal-Wallis). Spearman's test showed a perfect negative relationship (p=0.000, r= -0.975). Dunn's Post Hoc test was carried out as a follow-up test. Apoptosis results obtained p = 0.023 (Kruskal-Wallis). Spearman's test showed a perfect positive correlation (p=0.000, r=0.908). ANOVA test on BAX obtained significant differences between groups (p=0.000). The Tukey HSD test showed that there was a significant difference (p<0.05). The Pearson test showed a strong positive relationship between dose and BAX intensity (p=0.000, r-0.880). ANOVA test on Bcl-2 obtained a significant difference between groups (p=0.000). The Tukey HSD test showed a significant difference in the treatment group (p<0.05). The Pearson test indicated statistically that there was no significant relationship between dose and intensity of Bcl-2 with a weak positive correlation (p=0.435, r=0.249). Conclusion: C. racemosa extract can reduce viability, induce apoptosis, increase BAX expression. Increased expression of Bcl-2 in cell culture is a cell defense mechanism and is offset by increased expression of BAX so that cells continue to enter the apoptotic phase.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | 0423060084 |
| Subjects: | 600 Technology (Applied sciences) > 616 Diseases > 616.02 Special topics of disease > 616.025 Medical emergencies / Emergency medicine / Emergency nursing / Triage (Medicine) |
| Divisions: | S2/S3 > Magister Ilmu Biomedis, Fakultas Kedokteran |
| Depositing User: | Endang Susworini |
| Date Deposited: | 08 Dec 2023 08:59 |
| Last Modified: | 08 Dec 2023 08:59 |
| URI: | http://repository.ub.ac.id/id/eprint/205090 |
![[thumbnail of DALAM MASA MEMBARGO]](http://repository.ub.ac.id/style/images/fileicons/text.png) |
Text (DALAM MASA MEMBARGO)
NUR IEDHA TERTIANA.pdf Restricted to Registered users only until 31 December 2025. Download (4MB) |
Actions (login required)
 |
View Item |