毕业设计课题方向怎么写(开题范文)

调研结果1（文章的一部分）
• 样本：结直肠癌40例 • 方法：基因组DNA(FFPE)→杂交深度测序 (Agilent SureSelect custom
kit，包含14个重排相关基因（无NTRK1）) →Hiseq2000测序 • 结果：发现C2orf44-ALK融合 • 结论：影响临床靶向治疗的选择
CTB-35F21.1-PSD2
调研结果4
小结
• 结直肠癌中发现的融合位点有：
ALK
NTRK1 NTRK3 ROS RET VTI1A RSPO2 RSPO3 AKAP3
C2orf44；CAD; SPTBN1; EML4； PRKAR1A LMNA；TPM3 ETV6 SLC34A2 CCDC6；NCOA4 TCF7L2 EIF3E PTPRK RAD51AP1
3研究方案
筛选融合基因
已筛基因验证
下一步计 划
筛选融合基因
• Life Oncomine comprehensive assay • 样本：RNA
验证
• RT-PCR • Sanger sequensing • RACE技术
谢谢！
调研结果3-NTRK1融合治疗
调研结果4
• 样本：结直肠癌细胞系(SW620,LoVo, Co115, Colo320, IS1, IS2, IS3, TC7, TC71, FRI, V9P,LS174T, EB, and NCI-H508)
• RNA-seq筛选融合转录本 • RT-PCR及一代测序验证:细胞系 • 临床样本验证：106例结直肠癌 • 结果：发现三个新融合基因：AKAP13-PDE8A, COMMD10-AP3S1,
癌病人 • NTRK1融合治疗：细胞系、克唑替尼
调研结果3-NTRK1融合基因验证
调研结果3-NTRK1基因融合致瘤性分析
• In subsequent analyses of the oncogenicity of NTRK1 fusions, we generated LMNA(e6)-NTRK1(e11) or TPM3(e8)-NTRK1(e9, e11, e12) fusion transcripts harboring plasmid DNA for cell transformation. TrkA protein from a transformed NIH3T3 cell line with the fusion transcripts was well expressed (Figure 3A). The NIH3T3 cells overexpressing LMNA-NTRK1 (376 ± 33 colonies) or TPM3-NTRK1 (243 ± 46 colonies) formed a significantly larger number of colonies than nontransformed cells (1 ± 2 colonies, p < 0.01), which was to KM12 (285 ± 36 colonies) (Figure 3B). We then evaluated the tumorigenicity of NTRK1 fusions by inoculating immunocompromised athymic-mice with the transformed cells. Tumors from the transformed cells were palpable from day 18 of inoculation, and the volumes of the tumors were comparable to KM12 on day 29 of inoculation (Figure 3C). Although the expression level ofLMNA-TrkA fusion protein in NIH3T3 cells was less than that of the other two fusion protein-expressing cell lines, oncogenic activity was comparable with that of the other fusion proteins. Therefore, these results demonstrated that LMNA-NTRK1 and TPM3-NTRK1 could
调研结果1
调研结果2
• 样本：转移性结直肠癌 • 方法：DNA(FFPE) →靶向测序（19基因，Foundation Medicine）
→RET激酶抑制剂瑞戈非尼在RET融合阳性肿瘤细胞中的细胞毒作 用→RET融合阳性的结直肠癌患者治疗 • 结果：CCDC6-RET 和 NCOA4-RET发生融合
调研结果2
调研结果2
调研结果2
调研结果2
调研结果3
• 147结肠癌组织和47配对正常新鲜组织→RNA-seq→选择发现的 NTRK1融合基因进行验证（Sanger测序/FISH/IHC/）
• NTRK1融合致瘤性分析：从芯片、体内、体外三方面分析 • NTRK1融合临床意义：IHC 和FISH分析216 例韩国和472 中国结肠
RET激酶抑制剂只抑制RET融合阳性肿瘤细胞 患者CEA与LDH 下降
[1].
Le Rolle, A.F., et al., Identification and characterization of RET fusions in advanced colorectal cancer.
Oncotarget, 2015. 6(30): p. 28929-37.
[1].
Lipson, D., et al., Identification of new ALK and RET gene fusions from colorectal and lung cancer
biopsies. Nature Medicine, 2012. 18(3): p. 382-384.
结直肠癌融合基因的检测
内容
• 研究背景及意义 • 课题调研及结果 • 研究方案
Байду номын сангаас
研究背景及意义
• 结直肠癌是世界第三大恶性肿瘤，但只有一部融合基因在结直肠 癌中报道。
• 基因融合是由重排、倒位、串联重复等产生的 • 促进靶向药的研发；指导个体化治疗
课题调研
• CNKI • Pubmed • web of science • Google学术