Investigation on the function and mechanism of lincRNA TCONS_00024295 regulating CCR9 in T-lineage acute lymphocytic leukemia chemoresistance and metastasis
China (IFMSA-China) - Wuhan University School of Medicine, Wuhan
Xiaolian Zhang
Qiuping Zhang
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No No No No No No Yes No No No No
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy. The 5-year event-free survival remains at only 75 % and 50 % in children and adults, respectively. Chemoresistance and metastasis are the main factors threatening lives in leukemia patients. Accumulating evidences showed that high expression of CCR9 was closely associated with tumor chemoresistance and metastasis, and our early study indicated that the interaction of CL25/CCR9 involved in T-ALL chemoresistance and metastasis. LncRNA is a kind of noncoding RNA, which is more than 200nt and controls gene expression level on the transcriptional and post-transcriptional regulation. TCONS_00024295 is a new lncRNA screening by bioinformatics methods which highly correlates to the expression of CCR9, and it has not been reported before. Our research will explore the mechanism of TCONS_00024295 on CCR9 in T-ALL.TCONS_00024295 regulates CCR9 mediated T-ALL resistance
What is the aim of the project?
This study intends to explore whether TCONS_00024295 promotes CCR9 gene enhancer/promoter loop to regulate the expression of CCR9. Our research will shed novel insight on the prevention and clinical treatment of leukemia.
What techniques and methods are used?
1. Bioinformatics analysis about the CCR9 related lncRNA in T-ALL; 2. Expression levels of TCONS_00024295 in healthy donors’ and T-ALL patient’s peripheral blood; 3. Screened the most affected siRNA sequence of TCONS_00024295. 4. We use real-time PCR (Polymerase Chain Reaction) and Western-blot to detect the expression level of CCR9 and related Small interfering RNA (siRNA) in high-expression TCONS_00024295 cell line. Study on the effect of TCONS_00024295 on the regulation of CCR9.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Students will do: 1.The expression level of TCONS _ 00024295 in of 30 cases of T-ALL and 30 cases of normal controlled peripheral blood by real-time PCR; 2. RNA- fluorescence in situ hybridization (RNA-FISH) defines the expression level and location of TCONS _ 00024295 in T-ALL cell coatings slide; 3. The expression level of TCONS _ 00024295 in different T-ALL cell lines(MOLT-4, Jurkat, TALL-104, I2 .1, CCRF-CEM); 4. Select the high-expression TCONS _ 00024295 cell line, instantly reduce its expression, detected the expression level of CCR9 by Qpcr(Real-time Quantitative PCR Detecting System), WB(Western Blot) and flow cytometry. 5. Screened the most affected Small interfering RNA (siRNA) sequence of TCONS_00024295 by Bio-information assay. Then constructed for stable transfection cell lines.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary readings will be provided. In addition, the tutor will discuss and explain in detail about the project that students will be conducting. Students are encouraged to take part in the group meeting and seminars.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation
- The student will prepare a scientific report
- The student’s name will be mentioned in a future publication
- The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Cell culture, RNA and protein extraction, real-time PCR and Western-blotting
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- Students in biomedical fields
- Zhou B; Leng J; Hu M; Zhang L; Wang Z; Liu D; Tong X; Yu B; Hu Y; Deng C; Liu Y; Zhang Q*: Ezrin is a key molecule in the metastasis of MOLT-4 cells induced by CCL25/CCR9. Leuk Res 2010; 34(6):769-776.
- Zhang L; Xiao R; Xiong J; Leng J; Ehtisham A; Hu Y; Ding Q; Xu H; Liu S;Wang J; Tang DG; Zhang Q*: Activated ERM protein plays a critical role in drug resistance of MOLT-4 cells induced by CCL25. PLoS One 2013; 8(1): e52384.