CAJ | 학술논문

目的：研究外源性 DCC 基因稳定转染对人结直肠癌 SW1116细胞株的作用。方法采用反转录-聚合酶链反应从人正常结肠组织中扩增 DCC 基因功能区片段，构建 pcDNA3.1（+）-DCC 重组质粒并鉴定。将重组质粒转染入 DCC 基因缺失的结直肠癌 SW1116细胞中，用四甲基偶氮唑蓝（MTT）比色法观察其抑制结直肠癌细胞 SW1116的作用；用免疫荧光方法研究 pcDNA3.1（+）-DCC 质粒转染人结直肠癌细胞 SW1116后对结直肠癌细胞的影响及癌胚抗原（ CEA）表达。结果转染pcDNA3.1（+）-DCC 质粒的 SW1116细胞在转染后第3~6天，其细胞数显著低于转染 pcDNA3.1（+）质粒的 SW1116细胞和正常细胞对照（t =3.645，P ﹤0.05；t =3.132，P ﹤0.05）；转染 pcDNA3.1（+）-DCC 质粒的 SW1116细胞生长增殖速度低于转染 pcDNA3.1（+）质粒的 SW1116细胞和正常细胞对照（t =2.134，P ﹤0.05；t =2.736，P ﹤0.05）。转染 pcDNA3.1（+）-DCC 质粒的 SW1116细胞在转染后第2~6天，其总细胞活力显著低于正常细胞对照（t =3.053，P ﹤0.05）；转染 pcDNA3.1（+）-DCC 质粒的SW1116细胞在转染后第2、4、5、6天，其总细胞活力显著低于转染空质粒对照（t =2.816，P ﹤0.05）。转染 pcDNA3.1（+）-DCC 质粒的 SW1116细胞呈黄绿色荧光的细胞数量和荧光强度明显低于转染pcDNA3.1（+）质粒的 SW1116细胞和对照组细胞。结论转染 DCC 基因能抑制结直肠癌细胞的生长增殖，降低 CEA 表达，从而降低结直肠癌细胞浸润转移能力。
목적：연구외원성 DCC 기인은정전염대인결직장암 SW1116세포주적작용。방법채용반전록-취합매련반응종인정상결장조직중확증 DCC 기인공능구편단，구건 pcDNA3.1（+）-DCC 중조질립병감정。장중조질립전염입 DCC 기인결실적결직장암 SW1116세포중，용사갑기우담서람（MTT）비색법관찰기억제결직장암세포 SW1116적작용；용면역형광방법연구 pcDNA3.1（+）-DCC 질립전염인결직장암세포 SW1116후대결직장암세포적영향급암배항원（ CEA）표체。결과전염pcDNA3.1（+）-DCC 질립적 SW1116세포재전염후제3~6천，기세포수현저저우전염 pcDNA3.1（+）질립적 SW1116세포화정상세포대조（t =3.645，P ﹤0.05；t =3.132，P ﹤0.05）；전염 pcDNA3.1（+）-DCC 질립적 SW1116세포생장증식속도저우전염 pcDNA3.1（+）질립적 SW1116세포화정상세포대조（t =2.134，P ﹤0.05；t =2.736，P ﹤0.05）。전염 pcDNA3.1（+）-DCC 질립적 SW1116세포재전염후제2~6천，기총세포활력현저저우정상세포대조（t =3.053，P ﹤0.05）；전염 pcDNA3.1（+）-DCC 질립적SW1116세포재전염후제2、4、5、6천，기총세포활력현저저우전염공질립대조（t =2.816，P ﹤0.05）。전염 pcDNA3.1（+）-DCC 질립적 SW1116세포정황록색형광적세포수량화형광강도명현저우전염pcDNA3.1（+）질립적 SW1116세포화대조조세포。결론전염 DCC 기인능억제결직장암세포적생장증식，강저 CEA 표체，종이강저결직장암세포침윤전이능력。
Objective To investigate the effects of exogenous wild DCC gene stably transfection on growth of colorectal carcinoma cell line SW1116 in vitro. Methods DCC gene domain was amplified from human normal colon tissue by reverse transcript-polymerase chain reaction(RT-PCR). At first,a recombinant expression plasmid pcDNA3. 1( + )-DCC was constructed. Human colorectal carcinoma cell line SW1116 with-out DCC gene was transfected with pcDNA3. 1-DCC. Cell viability was tested by methyl thiazolyl tetrazolium (MTT)assay. Immunofluorescence staining was used to determine the effects of pcDNA3. 1-DCC and expres-sion of carcino-embryonic antigen(CEA)in human colorectal carcinoma cell line SW1116 which was transfect-ed with pcDNA3. 1-DCC. Results The population of cells transfected with pcDNA3. 1( + )-DCC plasmid was lower than those with pcDNA3. 1( + )-DCC plasmid and normal cells(t = 3. 645,P ﹤ 0. 05;t = 3. 132,P ﹤0. 05)at 3 ~ 6 days after transfection,and the proliferation rate of cells transfected with pcDNA3. 1( + )-DCC plasmid was lower than those with pcDNA3. 1( + )plasmid and normal cells(t = 2. 134,P ﹤ 0. 05;t = 2. 736, P ﹤ 0. 05). Cell line SW1116 transfected with pcDNA3. 1( + )-DCC plasmid total viability was lower than nor-mal cells(t = 3. 053 ,P ﹤ 0. 05)at 2 ~ 6 days after transfection. Cell line SW1116 transfected with pcDNA3. 1 ( + )-DCC plasmid total viability was lower than those with pcDNA3. 1( + )plasmid(t = 2. 816,P ﹤ 0. 05)at 2,4,5,6 days after transfection. The population of flavo-green colour cells transfected with pcDNA3. 1( + )-DCC plasmid and the fluorescent intensity of these cells were lower than those with pcDNA3. 1( + )plasmid and normal control cells. Conclusion Transfected DCC gene can suppress the cell proliferation and make CEA expression of cell line SW1116 down regulation to weaken its infiltration and metastasis abilities.