阿可的窩 Yi-Juai

Q: 為何做了 cDNA microarray, 還要做real-time PCR?

Apoptosis (Programmed Cell Death) Markers

(整理 by陳怡睿)

Loss of heterozygosity
(節錄自維基 by陳怡睿)

Gene silencing
(節錄自維基 by陳怡睿)

Epigenetics

(節錄自維基 by陳怡睿)

DNA methylation
(節錄自維基 by陳怡睿)

資料來源: 維基百科 整理: 陳怡睿

ELISA

ELISA (包括western blot及其比較與應用)

主題一: Stem Cell的定義

摘錄自: Stem Cell Information (The official National Institutes of Health resource for stem cell research)

Human papillomaviruses (HPVs) are small DNA tumor viruses that cause benign and malignant tumors of squamous epithelia.(Fig1) Among all the subtypes of HPVs, HPV-16 and HPV-18 are identified as high risk HPVs which are mainly responsible for HPV correlated human cancer. HPVs contain a double stranded DNA genome of approximately 8,000 base pairs and up to 10 open reading frames (ORFs).(Fig2) ORFs within the early region code for proteins like E6 and E7 which are involved in the regulation of viral replication and the viral life cycle, HPV-16 E6 and E7 oncoproteins can abrogate negative growth regulatory signaling pathways of the host cell through interaction with p53 and pRB tumor suppressor proteins. As a result, high-risk HPVs infected cells proliferation become de-regulated, and then, transformation develops.

聚合酵素鏈鎖反應(PCR)是一個簡便而有效的方法,它可使DNA在微量試管中擴增至106X以上。這個方法的原理十分簡單,在要擴增的DNA片段兩端分別設計一個前置引子(forward primer)和反置引子(reverse primer)使之與已變性的單股目標DNA緩冷配對(annealing)後,利用DNA聚合酵素(DNA polymerase)以目標DNA的兩股分別做為模板(template)來合成新的DNA股。像這樣經由(1)變性反應 (denaturation),使DNA的兩股分離。(2)緩冷配對反應(annealing)，使引子與目標DNA配對。(3)延長反應 (extension)，合成新的DNA股。的循環操作每次可使DNA的量添加一倍，若重複操作多次，以數學公式計算，DNA增加的量將會是2n，n是代表重複操作的次數。在理想的聚合酵素鏈鎖反應條件下，DNA是以幾何級數增加。理論上 ，一個DNA分子若重複操作 PCR 25次，那麼DNA的分子數將會擴增到225 = 106 個分子。這個DNA的量已足夠在Agarose凝膠電泳中觀察到。
PCR操作過程主要分成三大部份:(一)以高溫(92℃-95℃)使雙股模板DNA分離(denature)，(二)使引子與單股模板DNA做緩冷配對(40℃-52℃)，(三)再將溫度調整到DNA聚合酵素作用的有效溫度而合成新的DNA股。一般使用的DNA聚合酵素的有效作用溫度是37℃，因此在高溫分離雙股時會破壞DNA聚合酵素的活性，然而在耐高溫的細菌(Thermus aquaticus)中分離出來的DNA聚合酵素(Taq  DNA polymerase)在95℃中其活性的半衰期(half life)長達40分鐘，故可供PCR操作使用。Taq聚合酵素的有效作用溫度為72℃，在這溫度下，每分鐘可合成2000-4000個核甘酸(nucleotides)。由於Taq聚合酵素的發現，使PCR之操作得以自動化。

Genetic association of cytokine DNA polymorphisms with head and neck cancer
Oral Oncology (2008)

Pathogenesis of bone metastasis: a review

Ki-67 expression predicts locoregional recurrence in stage I oral tongue carcinoma
D Wangsa, M Ryott, E A° vall-Lundqvist, F Petersson, G Elmberger, J Luo, T Ried,

Involvement of NF-κB-mediated maturation of ADAM-17 in the invasion of oral squamous cell carcinoma
Yasuo Takamune, Tetsuro Ikebe, Osamu Nagano, Masanori Shinohara

ERBB receptors in developing, dysplastic and malignant oral epithelia
J. Rautava, K.J. Jee, P.J. Miettinen, B. Nagy, S. Myllykangas, E.W. Odell, T. Soukka, P.R. Morgan, K. Heikinheimo

DNA ploidy, proliferative capacity and intratumoral heterogeneity in primary and recurrent head and neck squamous cell carcinomas (HNSCC) – Potential implications for clinical management and treatment decisions
Holger G. Hass, Andreas Schmidt, Oliver Nehls, Stephan Kaiser

Clinicopathologic significance of EpCAM expression in squamous cell carcinoma of the tongue and its possibility as a potential target for tongue cancer gene therapy
EpCAM在舌鱗狀細胞癌的表達之臨床病理重要性以及其在舌癌基因治療中作為潛在標靶的可能性