CAJ | 학술논문

目的通过抑制转录共激活因子TAZ在结肠癌细胞中表达,探讨TAZ对结肠癌细胞的影响及机制.方法实时荧光定量聚合酶链反应(FQ-PCR)和Western blot方法检测TAZ在结肠癌组织和癌旁组织中表达的差异.合成针对TAZ的小干扰RNA(siRNA)转染结肠癌细胞株SW480,抑制TAZ表达;噻唑蓝(MTT)法检测细胞生长;细胞划痕实验检测细胞迁移活性;Transwell 小室侵袭实验检测细胞侵袭能力;FQ-PCR和Western blot检测增殖和迁移侵袭相关基因增殖细胞核抗原(PCNA)、细胞周期素(Cyclin) D1、p21、基质金属蛋白酶(MMP)-2、MMP-9、金属蛋白酶组织抑制因子(TIMP)-1的表达.结果结肠癌组织中TAZ mRNA(1.124±0.236)、蛋白表达(1.410±0.132)明显高于癌旁组织TAZ mRNA (0.428±0.163)、蛋白表达(0.559±0.089,P＜0.05).有效抑制SW480中TAZ表达后,转染组SW480细胞的活性在24 h(0.245±0.028)、48 h (0.413±0.051)、72 h(0.458 ±0.049)明显低于阴性对照组(分别为0.335±0.042、0.649±0.071、0.912±0.089);细胞划痕结果在TAZ-siRNA组、阴性对照组、空白组分别为17.17±1.96、38.67±5.88、42.20±4.34,Transwell小室实验结果分别为14.68±2.24、25.50±3.25、26.86±2.42,转染组的迁移和侵袭能力比阴性对照组和空白组明显减弱(P＜0.05).TAZ-siRNA转染后SW480中PCNA、Cyclin D1、MMP-1、MMP-2表达下调,而p21、TIMP-1的表达上调(P＜0.05).结论结肠癌细胞中的TAZ可以通过调节增殖和迁移侵袭相关基因而促进肿瘤细胞的生长、迁移和侵袭.
목적 통과억제전록공격활인자TAZ재결장암세포중표체,탐토TAZ대결장암세포적영향급궤제.방법 실시형광정량취합매련반응(FQ-PCR)화Western blot방법검측TAZ재결장암조직화암방조직중표체적차이.합성침대TAZ적소간우RNA(siRNA)전염결장암세포주SW480,억제TAZ표체;새서람(MTT)법검측세포생장;세포화흔실험검측세포천이활성;Transwell 소실침습실험검측세포침습능력;FQ-PCR화Western blot검측증식화천이침습상관기인증식세포핵항원(PCNA)、세포주기소(Cyclin) D1、p21、기질금속단백매(MMP)-2、MMP-9、금속단백매조직억제인자(TIMP)-1적표체.결과 결장암조직중TAZ mRNA(1.124±0.236)、단백표체(1.410±0.132)명현고우암방조직TAZ mRNA (0.428±0.163)、단백표체(0.559±0.089,P＜0.05).유효억제SW480중TAZ표체후,전염조SW480세포적활성재24 h(0.245±0.028)、48 h (0.413±0.051)、72 h(0.458 ±0.049)명현저우음성대조조(분별위0.335±0.042、0.649±0.071、0.912±0.089);세포화흔결과재TAZ-siRNA조、음성대조조、공백조분별위17.17±1.96、38.67±5.88、42.20±4.34,Transwell소실실험결과분별위14.68±2.24、25.50±3.25、26.86±2.42,전염조적천이화침습능력비음성대조조화공백조명현감약(P＜0.05).TAZ-siRNA전염후SW480중PCNA、Cyclin D1、MMP-1、MMP-2표체하조,이p21、TIMP-1적표체상조(P＜0.05).결론 결장암세포중적TAZ가이통과조절증식화천이침습상관기인이촉진종류세포적생장、천이화침습.
Objective The purpose of this study is to explore the effect and mechanism of TAZ to growth,migration and invasion in colon cancer cell line SW480 with RNA interference technology to inhibit expression of TAZ.Methods Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blot assays were used to detect the expression of TAZ in colon cancer tissues and normal tissues adjacent to cancer.Small interference RNA (siRNA) targeting TAZ were synthesized and transfected into SW480.Cell activity was measured with methyl thiazol tetrazolium (MTT) assay; Cell migration and invasion were observed via wound healing assay and transwell assay,respectively; The expressions of representative genes involved in proliferation and migration proliferating cell nuclear antigen (PCNA),Cyclin D1,p21,matrix metalloproteinase (MMP)-2,MMP-9,tissue inhibitorof metalloproteinase (TIMP)-1 were also detected by FQ-PCR and Western blotting assays.Results Expressions of TAZ mRNA (1.124 ± 0.236) and protein (1.410 ±0.132) in colon cancer tissues were significantly stronger than mRNA (0.428 ±0.163) and protein (0.559 ± 0.089)in normal tissues adjacent to cancer.After TAZ was suppressed,the cell activity of TAZ-siRNA group in 24 h (0.245 ±0.028),48 h (0.413 ±0.051),72 h (0.458 ±0.049) was weaker than that in control group (0.335 ± 0.042,0.649 ± 0.071,0.912 ± 0.089 respectively).Results of wound healing assay in TAZ-siRNA group,negative control group,blank control group were 17.17 ± 1.96,38.67 ± 5.88,42.20 ± 4.34; and results of transwell assay in 3 groups were 14.68 ± 2.24,25.50 ± 3.25,26.86 ± 2.42.And migration and invasion ability of TAZ-siRNA group were weaker than control groups (all P ＜ 0.05).In addition,the expression level of PCNA,Cyclin D1,MMP-2,MMP-9 decreased when TAZ was silenced,while the expression of P21,TIMP-1 increased.Conclusion TAZ in colon cancer cells can promote growth,migration and invasion ability of cancer cells by regulating proliferation,migration and invasion ralted genes.