癌症是全世界死亡的主要疾病，原因是患者在治療期間會產生抗藥性，導致腫瘤復發。腫瘤的異質性是抗藥的主要成因。如何在治療期間保持基因組的完整性將有利於提高生存率。在癌症進展中 p53 的突變與基因組不穩定性有關。雖然已經有許多抗藥性的研究在體外得到解決，但抗藥性到現在仍然是一個臨床未解決的問題。在這項研究中，使用體內動物模型來探討在有或沒有 TP53 的情況下 EGFR-TKI 誘導抗藥性的基因組完整性。本研究構建了 EGFRL858R 和 EGFRL858R x TP53+/- 兩品系小鼠的抗藥模型。於 H&E 染色結果，發現藥物敏感和腫瘤抗藥之間的細胞類型不同，在抗藥腫瘤中出現類似肌成纖維細胞的細胞型態，而在 TP53 突變的細胞核質比較大。其次，辨識藥物敏感、腫瘤抗藥和 TP53 突變小鼠的基因表達圖譜，發現三組基因組的突變熱點發生在染色體的不同區域。在腫瘤抗藥的情況下，TP53 突變以依賴性和非依賴性的方式發生不同突變。最後，所有有趣的突變基因也在 TCGA 數據庫中人類肺癌組進行了分析。未來，可利用西方墨點法和 IHC 測定小鼠和臨床樣品的蛋白表現。該研究不僅建立了體內抗藥動物，且釐清在抗藥產生的情形，基因體的穩定性以 p53 依賴性或非依賴性的方式產生的突變圖譜，這將有助於預防癌症治療過程中產生的抗藥性問題。

Cancer is a major cause of death worldwide. Many therapeutic strategies including surgery, chemotherapy, radiotherapy, target therapy and immune therapy have been developed up to now. However, drug resistance will be induced during the therapeutic period, subsequently leading recurrence. The heterogeneity of tumor is the major reason for inducing drug resistance. How to maintain the genomic integrity during therapeutic period will be beneficial for improving the survival rate. The p53 mutation occurred in cancer progression is involved in genomic instability. Although amount previous studies about drug resistance have been addressed in vitro, but this issue is still a clinical unmet up to now. In this study, we used in vivo animal model to explore the genomic integrity under TKI-induced drug resistance with or without TP53 haploinsufficiency. Two lines of mice, EGFRL858R and EGFRL858R x TP53+/-, have been constructed, thus induced cancer formation by doxycycline, subsequently treated with gefitinib for a long time until drug resistance. Several interesting findings have been found now. At first, the cellular types are very different between sensitive cancer and drug resistant cancer. One kind of myofibroblast-like cell was found in the lung tissue from drug resistant mice, not drug sensitive mice. Second, the gene expression profiles are distinguishable among sensitive, resistant and TP53 mutation mice. Third, the hotspots of drug-sensitive, drug resistance and loss of TP53 are induced in the different regions of chromosomes. Under drug resistant condition, chromosome 1 and chromosome 11 were highly mutation through the independent and dependent manner of TP53 mutation respectively. Finally, all the interesting mutation genes in the mice were also compared with the whole genome sequence of human lung cancer cohorts from TCGA. Data indicated that some genes are also mutated in clinical lung cancer patients. This study not only build up a in vivo drug resistant animal, but also provide the novel mutation profile in a TP53-dependent or independent manner during lung cancer progression and drug resistant period, which will contribute to the prevention of drug resistance during cancer therapy.

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