探討KIAA0101基因在肺癌中對生長與轉移所扮演的角色

KIAA0101基因在2001年被發現，它是一個proliferating cell nuclear antigen (PCNA)-associated factor，在KIAA0101基因第三段exon中的PCNA interaction protein box ( PIP box )會與p21競爭PCNA的結合位，進而促進細胞的增生，過去的文獻指出在肝臟、胃、肺等癌症中KIAA0101的高度表現可被當作是一個不好的預後因子，而近期的文獻中提到KIAA0101不僅能促進胃癌及腎上腺癌細胞的生長也能促進癌細胞的侵襲能力，因此本實驗將探討KIAA0101的表現對於肺癌細胞生長與轉移的影響，另外我們也將以PIP box mutant的KIAA0101來探討PIP box對於肺癌細胞的影響。在實驗中我們利用轉染技術將質體KIAA0101、與PIP box mutant的KIAA0101送入低表現的H1355與CL1-0肺癌細胞，在生長的實驗中提高KIAA0101的表現可以促進H1355肺癌細胞的生長並且與PIP box相關，但過度表現的KIAA0101卻無法促進CL1-0肺癌細胞的生長，而送入PIP box mutant的KIAA0101造成抑制細胞的生長，在轉移的實驗中發現提高KIAA0101的表現可以促進H1355及CL1-0肺癌細胞的爬行能力，而PIP box 的突變不影響KIAA0101促進H1355肺癌細胞的轉移能力，但其對CL1-0肺癌細胞的爬行能力卻會受到影響，我們以siRNA的方式抑制高表現KIAA0101的A549及CL1-5肺癌細胞，不論在生長或轉移的實驗中也都會受到影響，另外我們在明膠酶譜實驗中發現KIAA0101可以促進H1355肺癌細胞與侵襲能力有關的基質金屬蛋白酶MMP-2 & 9之活性，推測在肺癌細胞中KIAA0101與細胞也扮演促進侵襲能力之角色。

KIAA0101 gene was discovered in 2001, it is a PCNA associated factor, and was shown to bind PCNA competitively with the cell cycle regulator p21. PCNA is involved in DNA replication and damage repair. A number of studies have shown that overexpression of KIAA0101 predicts high stage, early tumor recurrence, and poor prognosis of many cancers and two recent studies has shown that overexpression of KIAA0101 promotes not only growth but also invasion in adrenal and gastric cancers, therefore, our studies is investigate the role of KIAA0101 gene in proliferation and metastasis of lung cancer. The other hand, we also use the plasmid which mutant the PIP box motif on KIAA0101 to investigate the function of PIP box motif of lung cancer. After plasmid transfection, the KIAA0101 is overexpression in H1355 and CL1-0 lung cancer cell. We using cell counting and colony formation assay to identify ability of growth. KIAA0101 promotes the growth in H1355 lung cancer cells, but it is not effect on CL1-0 lung cancer cells. The PIP box mutant KIAA0101 would inhibit cell growth in CL1-0 lung cancer cells. In migration and wound healing assays, KIAA0101 promotes migration in both of H1355 and CL1-0 lung cancer cells. After siRNA transfection, the expression of KIAA0101 is decreased in both of A549 and CL1-5 lung cancer cells. Our results shown that decrease expression of KIAA0101 reduce cell growth and migration both in A549 and CL1-5 lung cancer cells.