食道癌細胞にトランスポゾンを導入することで, 抗癌剤耐性遺伝子を増幅もしくは, 減弱させ, 抗癌剤暴露環境下で培養し, 作成した抗癌剤耐性株のDNA解析を行うことで, 耐性遺伝子を特定することが可能となる。食道癌細胞株TE4, 5, 9, 15について, Transposon tagged cellを樹立した。樹立したtransposon tagged cellを用いて, 各種薬剤を添加もしくは放射線照射を施行し, TE4, 15についてシスプラチン耐性株および放射線耐性株を獲得した。シスプラチン耐性遺伝子は37種類, 放射線耐性遺伝子は10遺伝子を同定され, このスクリーニングシステムが有用なことが証明された。
Transposon is a base sequence which transports to other location in the genome at random. Transposon is activated by transposase which is enzyme in transposon its self. Activatted transposon transports to other location in the genome (cut and paste). Inserting CMV promoter to transposon as a transcriptional activator, varioius cells which have vaious overexpressed genes or knocked down genes are obtained. Using this system into esophageal squamous cell cancer (ESCC), drug or radiation was added into these cells, and the resistant colonies were harvested to detect resistant gene. This new gene screening technique was useful for detecting candidate of drug-resistant genes and radio-resistant genes in esophageal squamous cell carcinoma. We identified 37 candidate genes responsible for CDDP resistance in the two cell lines derived from ESCC cells. Eleven genes were detected as candidate radioresistant genes in the two cell lines derived from ESCC cells.

研究種目 : 基盤研究(C)(一般)
研究期間 : 2013～2015
課題番号 : 25430159
研究分野 : 一般消化器外科