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发明公开
CN107142310A 特异性shRNA筛选及其靶向Ang‑2基因抑制肺癌细胞的验证方法 有权

特异性shRNA筛选及其靶向Ang‑2基因抑制肺癌细胞的验证方法
摘要：
本发明公开了特异性shRNA筛选及其靶向Ang‑2基因抑制肺癌细胞的验证方法，经细胞复苏、细胞培养、细胞转染、干扰前后实时荧光定量PCR检测Ang‑2表达、干扰前后细胞中蛋白表达分析、CCK‑8方法检测干扰Ang‑2基因前后的肺癌细胞增殖能力及干扰前后细胞侵袭、迁移能力改变的检测的步骤完成对特异性shRNA的筛选及其靶向Ang‑2基因抑制肺癌细胞的验证；本发明实现筛选对Ang‑2基因转录干扰具有特异性Ang‑2‑shRNA1有效质粒转染肺癌细胞，同时能够验证了特异性shRNA靶向干扰Ang‑2能够有效抑制癌细胞的增殖、侵袭、迁移等生物学能力，为治疗肺癌提供一种新的治疗方向，抑制Ang‑2弥补抗VEGFR单一疗法的局限性。
摘要（英）：

The invention discloses a method for specificity shRNA screening and verification on lung cancer cell inhibition of a targeted Ang-2 gene of shRNA. Specificity shRNA screening and verification on lung cancer cell inhibition of the targeted Ang-2 gene of the shRNA are completed through the steps of cell recovery, cell culture, cell transfection, Ang-2 expression detection through real-time fluorescent quantitative PCR before and after interference, expression analysis of protein in cells before and after interference, CCK-8 method based detection of lung cancer cell proliferation capability before and after Ang-2 gene interference and detection of cell invasion and migration capability change before and after interference. Screening of Ang-2-shRNA1 effective plasmid transfected lung cancer cells having specificity to Ang-2 gene transcription interference is achieved, meanwhile it can be verified that specificity shRNA targeted interference Ang-2 can effectively inhibit proliferation, invasion, migration and other biological capabilities of the cancer cells, a new treatment direction is provided for lung cancer treatment, the Ang-2 is inhibited, and the limitation of a single anti-VEGFR treating method is made up.

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