CAJ | 학술논문

目的：结肠癌的侵袭和转移是结肠癌患者死亡的主要原因。文中拟探讨叉头框蛋白M1（ Forkhead box M1， FoxM1）对人结肠癌细胞恶性表型的影响。方法应用实时荧光定量PCR（real time-PCR， RT-PCR）和Western blot方法筛选出高表达细胞株HT-29及低表达细胞株HCT-116，应用RNA干扰技术有效下调FoxM1在HT-29细胞中的表达，同时采用过表达质粒调高FoxM1在HCT-116中的表达，2种细胞株根据不转染、转染空载质粒及转染干扰或过表达 FoxM1质粒各分为3组：空白对照组、实验对照组、实验组。分别采用划痕实验和 Transwell小室法检测上述转染细胞的细胞增殖、迁移与侵袭能力的变化。结果 RT-PCR及Westen blot结果提示，FoxM1在HT-29细胞中高表达，而在HCT-116细胞中呈低表达。应用PEX-2-FoxM1上调HCT-116细胞中FoxM1表达后，细胞的划痕愈合能力HCT-116实验组[（70．92±1．48）％]较HCT-116实验对照组[（18．43±3．01）％]及HCT-116空白对照组[（16．66±2．63）％]显著增强（P ＜0．05）。 HCT-116实验组 Transwell 小室穿膜细胞数[（186．0±6．8）个]较HCT-116实验对照组[（42．0±2．0）个]及HCT-116空白对照组[（37．0±2．2）个]均增加（P＜0．05）。应用pGPH-sh FoxM1下调HT-29细胞中FoxM1表达后，HT-29实验组细胞的划痕愈合能力[（10．37±3．86）％]较HT-29实验对照组[（39．79±2．17）％]及HT-29空白对照组[（67．36±2．61）％]显著下降（P＜0．05）。 HT-29实验组Transwell 小室穿膜细胞数[（53．0±1．8）个]较较HT-29实验对照组[（95．0±2．2）个]及HT-29空白对照组[（118．0±4．0）个]均减少（P＜0．05）。结论FoxM1表达与结肠癌的侵袭、转移等关系密切；FoxM1的siRNA干扰可有效抑制结肠癌细胞的增殖、迁移和侵袭，人为调高结肠癌细胞株中FoxM1表达则促进细胞的增殖、迁移和侵袭。提示FoxM1可能成为抑制结肠癌细胞增殖和转移新的分子靶点。目的：結腸癌的侵襲和轉移是結腸癌患者死亡的主要原因。文中擬探討扠頭框蛋白M1（ Forkhead box M1， FoxM1）對人結腸癌細胞噁性錶型的影響。方法應用實時熒光定量PCR（real time-PCR， RT-PCR）和Western blot方法篩選齣高錶達細胞株HT-29及低錶達細胞株HCT-116，應用RNA榦擾技術有效下調FoxM1在HT-29細胞中的錶達，同時採用過錶達質粒調高FoxM1在HCT-116中的錶達，2種細胞株根據不轉染、轉染空載質粒及轉染榦擾或過錶達 FoxM1質粒各分為3組：空白對照組、實驗對照組、實驗組。分彆採用劃痕實驗和 Transwell小室法檢測上述轉染細胞的細胞增殖、遷移與侵襲能力的變化。結果 RT-PCR及Westen blot結果提示，FoxM1在HT-29細胞中高錶達，而在HCT-116細胞中呈低錶達。應用PEX-2-FoxM1上調HCT-116細胞中FoxM1錶達後，細胞的劃痕愈閤能力HCT-116實驗組[（70．92±1．48）％]較HCT-116實驗對照組[（18．43±3．01）％]及HCT-116空白對照組[（16．66±2．63）％]顯著增彊（P ＜0．05）。 HCT-116實驗組 Transwell 小室穿膜細胞數[（186．0±6．8）箇]較HCT-116實驗對照組[（42．0±2．0）箇]及HCT-116空白對照組[（37．0±2．2）箇]均增加（P＜0．05）。應用pGPH-sh FoxM1下調HT-29細胞中FoxM1錶達後，HT-29實驗組細胞的劃痕愈閤能力[（10．37±3．86）％]較HT-29實驗對照組[（39．79±2．17）％]及HT-29空白對照組[（67．36±2．61）％]顯著下降（P＜0．05）。 HT-29實驗組Transwell 小室穿膜細胞數[（53．0±1．8）箇]較較HT-29實驗對照組[（95．0±2．2）箇]及HT-29空白對照組[（118．0±4．0）箇]均減少（P＜0．05）。結論FoxM1錶達與結腸癌的侵襲、轉移等關繫密切；FoxM1的siRNA榦擾可有效抑製結腸癌細胞的增殖、遷移和侵襲，人為調高結腸癌細胞株中FoxM1錶達則促進細胞的增殖、遷移和侵襲。提示FoxM1可能成為抑製結腸癌細胞增殖和轉移新的分子靶點。
목적：결장암적침습화전이시결장암환자사망적주요원인。문중의탐토차두광단백M1（ Forkhead box M1， FoxM1）대인결장암세포악성표형적영향。방법응용실시형광정량PCR（real time-PCR， RT-PCR）화Western blot방법사선출고표체세포주HT-29급저표체세포주HCT-116，응용RNA간우기술유효하조FoxM1재HT-29세포중적표체，동시채용과표체질립조고FoxM1재HCT-116중적표체，2충세포주근거불전염、전염공재질립급전염간우혹과표체 FoxM1질립각분위3조：공백대조조、실험대조조、실험조。분별채용화흔실험화 Transwell소실법검측상술전염세포적세포증식、천이여침습능력적변화。결과 RT-PCR급Westen blot결과제시，FoxM1재HT-29세포중고표체，이재HCT-116세포중정저표체。응용PEX-2-FoxM1상조HCT-116세포중FoxM1표체후，세포적화흔유합능력HCT-116실험조[（70．92±1．48）％]교HCT-116실험대조조[（18．43±3．01）％]급HCT-116공백대조조[（16．66±2．63）％]현저증강（P ＜0．05）。 HCT-116실험조 Transwell 소실천막세포수[（186．0±6．8）개]교HCT-116실험대조조[（42．0±2．0）개]급HCT-116공백대조조[（37．0±2．2）개]균증가（P＜0．05）。응용pGPH-sh FoxM1하조HT-29세포중FoxM1표체후，HT-29실험조세포적화흔유합능력[（10．37±3．86）％]교HT-29실험대조조[（39．79±2．17）％]급HT-29공백대조조[（67．36±2．61）％]현저하강（P＜0．05）。 HT-29실험조Transwell 소실천막세포수[（53．0±1．8）개]교교HT-29실험대조조[（95．0±2．2）개]급HT-29공백대조조[（118．0±4．0）개]균감소（P＜0．05）。결론FoxM1표체여결장암적침습、전이등관계밀절；FoxM1적siRNA간우가유효억제결장암세포적증식、천이화침습，인위조고결장암세포주중FoxM1표체칙촉진세포적증식、천이화침습。제시FoxM1가능성위억제결장암세포증식화전이신적분자파점。
Objective The invasion and metastasis of colon cancer often leads to treatment failure and mortality in patients . Our research is to investigate the influence of FoxM 1 to malignant human colon cancer line . Methods In two human colon cancer lines, the protein and mRNA expression levels of FoxM 1 were analyzed with the application of RT-PCR and Western blot , from which high-expressed HT-29 and low-expressed HCT-116 were determined.The expression of FoxM1 was down-regulated by RNA interfering in HT-29 and up-regulated by constructing overexpression transgenic line in HCT-116.The proliferation of the above cells was assayed by healing method;while the metastasis and invasion ability were examined by Transwell chamber assay . Results Two colon cancer lines were selected with high-expression or low-expression of FoxM1 separately named HT-29 and HCT-116.Application of PEX-2-FoxM1 raised after HCT-116 cells express FoxM1, cell scratches in HCT-116 experimetal group ([70.92 ±1.48]%) compared with HCT-116 control group([16.92 ±4.05]%)and HCT-116 blank control group([16.66 ±2.63]%) will markedly enhance its capabil-ity of healing (P<0.05), Transwell Chambers in membrane cells in HCT-116 experimetal group (186.0 ±6.8) compared with HCT-116 control group(42.0 ±2.0) and HCT-116 blank control grou (37.0 ± 2.2)was increased (P<0.05).On the other hand, the applied pG-PH-shFoxM1 can reduce FoxM1 expression in HT-29 cell, cell scrat-ches healing ability in HT-29 experimetal group ( [ 10 .37 ± 3.86]%) compared with HT-29 control group([34.63 ±2.35]%)and HT-29 blank control group([67.36 ±2.61]%) decreased significantly (P<0.05), Transwell Chambers in membrane cells in HT-29 experimetal group (53.0 ±1.8)compared with HT-29 control group(95.0 ±2.2)and HT-29 blank control grou(118.0 ±4.0) was also reduced (P<0.05). Conclusion The expression of FoxM1 is in close relation to the invasion and metastasis of CRC .The fact that the siRNA interfering FoxM1 could effectively inhibit the proliferation, metastasis and invasion, suggesting FoxM1 could po-tentially be a new molecular target for inhibiting the proliferation of human colon cancer line .