二、細胞實驗效果不錯,那動物實驗怎么做?
動物模型可在在腫瘤基礎性研究和抗腫瘤研究中發揮重要作用。結直腸癌研究中動物實驗主要分為三種類型:I自發結直腸癌模型;II誘導結直腸癌模型;III 移植結直腸癌模型。第I種主要是通過構建轉基因動物的方式實現,第II種則利用化學誘導劑誘導結直腸癌發生,但是誘導模型不便于研究腫瘤的侵襲和轉移能力;第III種將人類腫瘤組織或細胞直接接種于裸鼠進行研究。因裸鼠培育條件低,無毛,易于動態觀察腫瘤的生長狀態,且通過接種方式、部位的選擇可達到較滿意的轉移效果,因此目前多選擇裸鼠進行人結直腸癌細胞異體移植。
據數據統計,約30%的CRC患者發生肝臟轉移,這也是造成CRC患者死亡的最主要原因。今天我們就來著重看一看,結直腸癌轉移模型的構建吧。
注:增殖方向體內模型為裸鼠皮下成瘤,屬于腫瘤研究的常用模型,今天就不做詳細介紹了。實驗方法可參考前兩期肺癌、胃癌的總結文章。
三、體內外模型選好了,那么我的實驗需要使用哪種慢病載體對基因進行操作呢?
啟動子、熒光標記、抗性標簽是我們選擇載體需要考慮的三大要素。實驗不同,這些元件也需要相應變化。對于腸癌研究來說,人腸上皮細胞表面抗原基因A33啟動子,可作為組織特異性啟動子使用。按例祭出神圖一張,定制你的專屬載體,就是這么簡單!

參考文獻
【1】 Ahmed, D., et al. “Epigenetic and genetic features of 24 colon cancer cell lines.” Oncogenesis 2.9 (2013): e71.
【2】 Messner, Isabelle, et al. “KRAS p. G13D mutations are associated with sensitivity to anti-EGFR antibody treatment in colorectal cancer cell lines.”Journal of cancer research and clinical oncology 139.2 (2013): 201-209.
【3】 Ji-Fu, E., et al. "Suppression of lung cancer metastasis-related protein 1 (LCMR1) inhibits the growth of colorectal cancer cells." Molecular biology reports 39.4 (2012): 3675-3681.
【4】 Xing, Cheng, et al. “Ubiquitin-specific protease 4-mediated deubiquitination and stabilization of PRL-3 is required for potentiating colorectal oncogenesis.” Cancer research 76.1 (2016): 83-95.
【5】 Yuan, Wei, et al. "Up-regulation of microRNA-145 associates with lymph node metastasis in colorectal cancer." PloS one 9.7 (2014): e102017.
【6】 Zhuang, Kangmin, et al. “CDK5 functions as a tumor promoter in human colorectal cancer via modulating the ERK5–AP-1 axis.” Cell Death & Disease 7.10 (2016): e2415.
【7】 He, Xiao-Wen, et al. "NOB1 is essential for the survival of RKO colorectal cancer cells." World Journal of Gastroenterology: WJG 21.3 (2015): 868.
【8】 Wang, Xiao, et al. “Metalloproteases meprin-ɑ (MEP1A) is a prognostic biomarker and promotes proliferation and invasion of colorectal cancer.”BMC cancer 16.1 (2016): 383.
【9】 Ji, Dengbo, et al. "MicroRNA-181a promotes tumor growth and liver metastasis in colorectal cancer by targeting the tumor suppressor WIF-1." Molecular cancer 13.1 (2014): 1.
【10】 Huang, Huang, et al. “Regulation of TWIK-related potassium channel-1 (Trek1) restitutes intestinal epithelial barrier function.” Cellular & molecular immunology 13.1 (2016): 110-118.
【11】 Leng, Zhengwei, et al. “Krüppel-like factor 4 acts as an oncogene in colon cancer stem cell-enriched spheroid cells.” PloS one 8.2 (2013): e56082.
【12】 Urosevic, Jelena, et al. "Colon cancer cells colonize the lung from established liver metastases through p38 MAPK signalling and PTHLH." Nature cell biology 16.7 (2014): 685-694.
【13】 Okamoto, Koji, et al. “miR‐493 induction during carcinogenesis blocks metastatic settlement of colon cancer cells in liver.” The EMBO journal 31.7 (2012): 1752-1763.