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The role of kinesin family member 16B in tumorigenesis

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Author(s)
김별
Issued Date
2020
Abstract
종양은 정상세포가 오랜기간 돌연변이가 축적되어 형질전환 된 후 계속적인 분열이 발생되고 이동이 가능한 형태로 진행된다. 많은 유전자에서 돌연변이가 발생되는데 아직도 정확한 원인 유전자들이 완전하게 규명되지 않았다. 따라서 종양 진단과 타겟치료제 개발에 현재까지 많은 한계점을 지니고 있다.
새로운 차료타겟을 찾아내기 위해 정상세포와 종양세포에서 그 발현이 현저하게 차이가 있는 유전자를 데이터베이스에서 바이오인포메틱 기법을 통하여 kinesin family member 16B (KIF16B)를 발견하였다.
이 연구에서는 정상과 종양 암 환자 조직에서 그 발현이 차이가 있는 KIF16B를 통해 이 유전자가 종양형성에 어떠한 영향을 끼치는지를 관찰하고자 하였다. 우리는 폐 암 세포에서의 KIF16B를 konckdown을 통해 mRNA와 protein의 발현이 감소하는 것을 확인하였다. 또한 MTT assay와 Soft agar colony formation assay를 통해 세포의 성장과 증식의 감소를 확인하였고, Transwell migration assay를 통해 세포의 이동성이 감소하는 것을 확인하였다. 이러한 oncogenic effect는 KIF16B가 knockdown 된 폐 암 세포에서 in vitro 연구를 통해 현저하게 감소되었다. 이와 같은 결과는 KIF16B가 in vitro assay에서 암 세포의 종양 형성 가능성과 관련 될 수 있음을 보여준다. 따라서, 이 연구결과들을 통해 향 후 KIF16B를 이용한 종양 진단 및 치료제 개발에 중요한 정보를 제공할 것으로 판단된다. | Cancer are formed in such a way that normal cells undergo transformation and accumulate long-term mutations, and then undergo continuous division and migration. Mutations occur in many genes, but the exact causal genes are not fully identified. Therefore, there are many limitations to the present diagnosis of tumor diagnosis and the development of target therapies. In order to find new car targets, kinesin family member 16B (KIF16B) was found in the database using genes with markedly different expressions in normal and tumor cells. In this study, we examined the effect of this gene on tumorigenesis through KIF16B, whose expression differs in normal and tumor cancer tissues. Here, we show that KIF16B knockdown decrease mRNA and protein expression. In addition, cell growth and proliferation were reduced by MTT assay and soft agar colony for-mation assay, and cell mobility was decreased by Transwell migration assay. These oncogenic effects were significantly decreased in KIF16B knockdown lung cancer cells through in vitro studies. These results demonstrated that KIF16B could be associated with tumorigenic potential of cancer cells in vitro assays. Taken together, these results will provide important information for the future diagnosis and treatment of tumors using KIF16B.
Alternative Title
The role of kinesin family member 16B in tumorigenesis
Alternative Author(s)
Byeol Kim
Department
일반대학원 의과학과
Advisor
유호진
Awarded Date
2020-02
Table Of Contents
KOREAN ABSTRACT…………………………………………………………Ⅴ
INTRODUCTION……………………………………………………………………1
MATERIALS AND METHODS
1. Cell culture and treatment………………………………………………………………6
2. Small interfering RNA for KIF16B and cell transfection……………………………………………………………………………………7
3. RNA isolation and cDNA synthesis……………………………………………………8
4. Quantitative real-time PCR………………………………………………………………9
5. Western blot analysis……………………………………………………………………10
6. MTT assay…………………………………………………………………………………11
7. Cell migration assay…………………………………………………………………… 12
8. Soft agar assay……………………………………………………………………………13
9. FACS analysis……………………………………………………………………………13
10. Statistical analysis………………………………………………………………………14
RESULTS
1. The expression of KIF16B upregulated in lung cancer…………………………15
2. KIF16B expression inhibits by small-interference(siRNA) targeting KIF16B…………………………………………………………………………………………18
3. Knockdown of KIF16B inhibited proliferation ability in lung cancer cells………………………………………………………………………………………………21
4.Knockdown of KIF16B arrested cell cycle in the G2/M phase……………………………………………………………………………………………24
5. Knockdown of KIF16B suppresses migration ability in lung cancer cells ……………………………………………………………………………………………………29
DISCUSSION
Discussion……………………………………………………………………………………32
ABSTRACT
Abstract……………………………………………………………………………………37
REFERENCES
References………………………………………………………………………………39
Degree
Master
Publisher
조선대학교 대학원
Citation
김별. (2020). The role of kinesin family member 16B in tumorigenesis.
Type
Dissertation
URI
https://oak.chosun.ac.kr/handle/2020.oak/14072
http://chosun.dcollection.net/common/orgView/200000278450
Appears in Collections:
General Graduate School > 3. Theses(Master)
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