Ph.D. by Research at UNPAD : Potential Effect of Cigarrette on promoter p16 in CCL-171 cell

Information: This scholarship is funded by Universitas Padjadjaran - Indonesia.

Subject Areas: Pharmaceutical Sciences

Project Title: Potential Effect of e-Cigarrete, Ciggarete, Cigaratte Reference stimulates hypermethylation promoter p16 and its correlation with characteristic changes in CCL-171 cell line

Project Supervisor: Melisa Intan Barliana, Dr. Med. Sc., Apt.

Project Description:

Smoke exposure of cigarette has been shown to cause development and progression of lung cancer. Currently, cigarette products are not only conventional cigarettes (with burning) but electronic nicotine delivery systems (ENDs) or e-Cigs and e-liquids. This product increases the risk of developing and progressing lung disease even though conventional cigarettes have a higher risk. Conventional cigarettes have been shown to hypermethylate several promoters resulting in transcriptional silencing of several suppressor genes, such as genes involved in the cell cycle (p16), in apoptosis (DAPK), and in proliferation and differentiation (RARβ). The prevalence of lung cancer may affected by inter-individual character such as genetic polymorphisms. For example genes involved in the metabolism of carcinogenic compounds found in cigarettes (CYP1A1) and genes involved in detoxification (GSTM1). Thus, the aim of this study is to analyze the risk progression of lung cancer in normal lung cells due to the effect of ENDs on the incidence of hypermetylation of the promoter p16, DAPK, and RARβ, as well as changes in characteristic of the cells. Furthermore, to analyze how the effect of CYP1A1 and GSTM1 gene polymorphisms on the hypermetylation. The inflammatory response and antioxidant status will also be analyzed after exposure to cigarette smoke. Exposure to cigarette smoke will use smoking machines LM1 and LM4 (Borgwaldt) on normal lung cell culture (CCL-171). The smoke of e-Cigarettes (e-Cigs), cigarettes (conventional cigarettes), and reference cigarettes (1R6F) will be exposed to normal lung cells CCL-171. In the first year, there will be cytotoxicity, promoter methylation, and cell morphology analysis. Methylation analysis of the promoter gene will use the Methylation Specific PCR (MSP) method with specific primers for p16. Cell morphology will be analyzed using Odissey LiCOR. In the second year, analysis of cytokines, antioxidant status, and gene construction will be carried out. The cytokines to be analyzed were IL-6 and IL-8 using the ELISA method. Measurement of antioxidant status through measurement of total glutathione (GSH) using a colorimetric method. Next, the construction of CYP1A1*2A and GSTM1 genes will be carried out which will be confirmed by sequencing. The third year will carry out the transfection of constructed-genes that to normal lung cells. Furthermore, cigarette smoke exposure will be exposed to normal lung cells that had been transfected with the CYP1A1*2A or GSTM1 gene, and MSP analysis will be done on p16, and cell morphology. This study will confirm smoke exposure to normal lung cells affecting lung cancer development in vitro.

1. This scholarship is open to candidates of any nationality;
2. Applicants should hold a minimum of Master's degree (or its equivalent) in Pharmaceutical Sciences. However, other related discipline with relevant experience will be considered.;
3. Age limit: Applicants must be under 31 years old;
4. Minimum GPA of 3.5 (out of 4) or equivalent. Academic transcripts are required from every college you have attended for at least one academic year as a full-time student. Your transcripts should be uploaded as a scanned copy or PDF. Please ensure that the degree conferral date and the degree conferred is clearly visible on the document.
5. English Language Requirements:
6. You need to achieve the minimum scores shown for any of the English language tests below:

• Minimum overall score for International TOEFL (Paper-Based Test): 525 OR;
• Minimum overall score for International TOEFL (Internet-Based Test): 70 OR;
• Minimum overall score for International TOEFL (Computer-Based Test): 190 OR;
• Minimum overall score for International English Language Testing System (IELTS) Academic: 6.

1. Full cost of tuition fees at the Doctoral Program in Universitas Padjadjaran;
2. Monthly living allowance IDR 2.500.000 for 3 (three) years during the Doctoral Program;
3. 1 (one) time return airfare ticket to and from Jakarta Indonesia (from and to the candidate's nearest international airport);
4. Health insurance for 3 (three) years, during the Doctoral Program;
5. There will be additional funds available for research expenses for 3 (three) years during the doctoral program.

To apply, please complete and submit the following documents to international@unpad.ac.id, and address the email for your application as 'Padjadjaran Doctoral Scholarship – Melisa Intan Barliana':

1. Degree certificates and transcripts;
2. Two letter of recommendations. Applicants are required to submit two letters of recommendation. These recommendations should be written by people who have supervised you in an academic, employment, or community service setting. Your recommendations should directly address your suitability for admission to a postgraduate program at Universitas Padjadjaran;
3. Statement of Purpose. Your statement should be typed, single-spaced and should be between two to three pages. Describe your reasons for applying to the proposed program, your preparation for this field of study, and explain why the position particularly matches your skills and experience, and how you choose to develop the project, which may aid the admissions committee in evaluating your motivation for postgraduate study.
4. 2-page Curriculum Vitae;
5. Certificate of Proficiency in English; and
6. Other supporting documents.

If you require any further information with regard to this, please contact international@unpad.ac.id.