目的糖尿病患者结肠癌的发病率显著高于非糖尿病人群,晚期糖基化终产物(advanced glycation end products,AGEs)在糖尿病患者体内生成明显增多。文中观察AGEs对人结肠癌细胞SW-480增殖的影响及其机制。方法体外培养人结肠癌细胞SW-480,...目的糖尿病患者结肠癌的发病率显著高于非糖尿病人群,晚期糖基化终产物(advanced glycation end products,AGEs)在糖尿病患者体内生成明显增多。文中观察AGEs对人结肠癌细胞SW-480增殖的影响及其机制。方法体外培养人结肠癌细胞SW-480,以终浓度分别为100μg/ml、200μg/ml、500μg/ml的AGE-BSA处理细胞24h,并设正常对照组和BSA对照组进行比较。采用四甲基偶氮唑盐(MTT)法检测SW-480细胞活力,利用流式细胞术检测细胞周期的改变,实时荧光定量PCR测定晚期糖基化终产物受体(receptor for advanced glycation end products,RAGE)mRNA、细胞周期素D1(cyclin D1)mRNA的表达。结果①MTT结果示100μg/ml、200μg/ml、500μg/mlAGE-BSA作用SW-480细胞24 h后可以显著促进细胞增殖(P<0.05),并呈浓度依赖性。②流式细胞术结果示200μg/mlAGE-BSA干预组24 h后可以减少SW-480细胞G1期百分率,同时增加S期百分率,与正常对照组比较具有统计学差异(P<0.05)。③实时荧光定量PCR结果示与正常对照组相比,200μg/mlAGE-BSA干预后可以上调RAGEmRNA和cyclin D1 mRNA的表达(P<0.05)。结论 AGEs能促进SW-480细胞的增殖,其机制可能与上调RAGEmRNA和cyclin D1 mRNA的表达,加速细胞G1期向S期转换相关。展开更多
Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically i...Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically inactive mutant forms of those enzymes in SW620 and SW480 cells, two human cell lines derived from non-metastatic cancer and metastatic colorectal cancer, respectively. While the expression of the native forms of PRLs promotes the spreading, adhesion, and proliferation of these cells, the expression of their mutant forms inhibits the earlier-mentioned processes. These data thus provide a cellular mechanism for the role of PRL-3 in tumor metastasis and suggest that all the three PRLs have similar functions.展开更多
In order to examine the effect of GRIM19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIM19 cDNA was amplified by PCR with the template...In order to examine the effect of GRIM19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIM19 cDNA was amplified by PCR with the template pcxn2-GRIM19 and cloned into the shuttle plasmid pAdTrack-CMV. The plasmid pAdTrack-CMV-GRIM19 was linearized by PmeⅠ and homologously recombined with bone plasmid pAdEasy-1 in BJ5183, followed by identification by enzyme digestion. After transfection of linearized pAd-GRIM19 with PacⅠ into HEK293 cells, Ad-GRIM19 was obtained and amplified by 3 circles. SW480 cells were infected with Ad-GRIM19. The apoptosis rate was detected by flow cytometry. Agarose electrophoresis revealed the bands of recombinant plasmids identified by enzyme digestion were in the right range corresponding with expectation. Under the fluorescent microscopy, the package of Ad-GRIM19 in HEK293 cells and the expression of Ad-GRIM19 in SW480 cells were observed. The transfection of Ad-GRIM19 into SW480 cells increased the apoptosis rate of SW480 cells as compared with controls, It was concluded that Ad-GRIM19 was successfully constructed and the overexpression of GRIM19 in colon cancer cell lines could promote the apoptotic cell death.展开更多
Objective： To investigate the effect of DL-lysine acetylsalicylate on proliferation of colon carcinoma cells line sw480. Methods： After treatment of DL-lysine acetylsalicylate, the study was performed by observing s...Objective： To investigate the effect of DL-lysine acetylsalicylate on proliferation of colon carcinoma cells line sw480. Methods： After treatment of DL-lysine acetylsalicylate, the study was performed by observing sw480 colorectal cancer cells with phase contrast microscope, making growth curve, and examining the inhibition rate of sw480 cells with MTT assay. Results： The morphology of sw480 cells showed characteristics of apoptosis, the cell growth curve showed inhibited proliferation of sw480 cells when treated with DL-lysine acetylsalicylate （P〈0.05）. The rate of inhibition was upward when the drug concentration increased. Conclusion： DL-lysine acetylsalicylate for injection can inhibit the growth of sw480 colorectal cancer cells obviously in a dose dependent manner.展开更多
[Objectives] The aim was to elucidate the pro-apoptosis mechanism of naphthoquinone derivative 2-octyl sulfoxide-1,4-naphthoquinone(OSNQ) on human colon cancer SW480 cells.[Methods]The cytotoxic effect of OSNQ on colo...[Objectives] The aim was to elucidate the pro-apoptosis mechanism of naphthoquinone derivative 2-octyl sulfoxide-1,4-naphthoquinone(OSNQ) on human colon cancer SW480 cells.[Methods]The cytotoxic effect of OSNQ on colon cancer SW480 cells was detected by MTT colorimetry.The pro-apoptotic effect of OSNQ on human colon cancer SW480 cells was detected by Annexin V-FITC/PI double staining.The changes in expression of apoptosis-related proteins were detected by Western blot.[Results]The results of MTT assay showed that OSNQ had a significant cytotoxic effect on colon cancer SW480 cells.The results of Western blot showed that OSNQ induced the apoptosis in colon cancer SW480 cells through promoting the expression of pro-apoptotic caspase-3 and inhibiting the expression of apoptosis-inhibiting protein Bcl-2.[Conclusions] OSNQ has a significant cytotoxic effect on colon cancer SW480 cells,and it induces the apoptosis of colon cancer SW480 cells by AKT signaling pathway.展开更多
目的观察青藤碱对高侵袭性人结肠癌细胞株SW 480移植瘤细胞周期及结肠瘤体组织中环氧合酶-2(COX-2)表达的影响,研究青藤碱防治结肠癌的机制。方法先用5只免疫功能正常的BALB/c-nu/nu裸小鼠建立高侵袭性人结肠癌瘤源,3周后取直径约2 mm...目的观察青藤碱对高侵袭性人结肠癌细胞株SW 480移植瘤细胞周期及结肠瘤体组织中环氧合酶-2(COX-2)表达的影响,研究青藤碱防治结肠癌的机制。方法先用5只免疫功能正常的BALB/c-nu/nu裸小鼠建立高侵袭性人结肠癌瘤源,3周后取直径约2 mm的瘤块移植于20只免疫缺陷的BALB/c-nu/nu2裸小鼠回盲浆膜凹龛部,1周后再随机分为结肠癌组和青藤碱组。青藤碱组按10 m L/(kg·d)灌10%青藤碱治疗30 d,结肠癌组灌0.9%氯化钠注射液对照。测量BALB/c-nu/nu裸小鼠结肠瘤体积、瘤质量并计算肿瘤抑制率;流式细胞仪检测结肠瘤体组织SW 480细胞在G1、G2、M和S期的细胞周期比例;免疫组化法检测结肠瘤体组织COX-2的表达,RT-PCR法检测COX-2 m RNA表达。结果与结肠癌组比较,青藤碱组结肠瘤体积、瘤质量及S期细胞比值明显降低、G1、G2期细胞比值提高,肿瘤抑制率为(39.75%)(P<0.05);COX-2和COX-2 m RNA低表达(P<0.05)。结论青藤碱可能在G1、G2和S期诱导SW 480细胞阻滞、抑制结肠癌细胞株SW 480细胞的有丝分裂,并通过下调COX-2表达而间接影响癌细胞的增殖,具有抗高侵袭性人结肠癌的作用。展开更多
文摘目的:探讨上皮膜蛋白1(epithelial membrane protejn-1,EMP1)在人结直肠癌(colorectal carcinoma,CRC)组织中的表达水平及其过表达对CRC SW-480细胞增殖、凋亡和侵袭的影响。方法:免疫组织化学、Western blotting方法检测63例CRC组织及31例癌旁组织中EMP1的表达水平,分析EMP1表达与CRC临床病理参数的关系。慢病毒介导将pLenti6-EMP1质粒转染SW-480细胞,建立EMP1过表达细胞,转染plenti6/V5-DEST空白质粒为对照。Real-time PCR及Western blotting检测转染后SW-480细胞株中EMP1的表达,MTT、流式细胞术及Transwell实验分别检测EMP1过表达对SW-480细胞增殖、凋亡及侵袭能力的影响。结果:EMP1蛋白在人CRC组织的表达显著低于癌旁组织(0.257±0.022 vs 0.863±0.086,P<0.05),并且其表达水平在不同T分期、有无淋巴结转移、临床分期以及组织分级组间表达有显著差异(P<0.05),而与患者年龄、性别、肿瘤部位无关(P>0.05)。成功构建EMP1过表达的LeEMP1细胞。与LeEmpty细胞相比,LeEMP1细胞的增殖能力显著下降[(60.94±4.04)%vs(100.00±0.00)%,P<0.05],凋亡率显著升高[(12.10±1.30)%vs(3.10±0.60)%,P<0.05]、侵袭转移能力显著降低[穿膜细胞数:(87.00±12.00)vs(178.00±21.00)个,P<0.05]。LeEMP1细胞Caspase-9表达显著高于LeEmpty细胞(0.764±0.073 vs 0.231±0.029,P<0.05),VEGFC表达显著降低(0.185±0.022 vs 0.663±0.065,P<0.05)。结论:人CRC组织中EMP1蛋白表达明显减低,过表达EMP1能够抑制CRC SW-480细胞的恶性生物学行为,其可能通过调控Caspase-9和VEGFC的表达来发挥作用。
文摘Recent studies suggest that PRL-3 is involved in the metastasis of colorectal cancer, but the mechanism concerning that has not been well defined. This article expresses PRL-1, PRL-2, and PRL-3 and the catalytically inactive mutant forms of those enzymes in SW620 and SW480 cells, two human cell lines derived from non-metastatic cancer and metastatic colorectal cancer, respectively. While the expression of the native forms of PRLs promotes the spreading, adhesion, and proliferation of these cells, the expression of their mutant forms inhibits the earlier-mentioned processes. These data thus provide a cellular mechanism for the role of PRL-3 in tumor metastasis and suggest that all the three PRLs have similar functions.
基金a grant from National Natu-ral Sciences Foundation of China (No. 30471673)
文摘In order to examine the effect of GRIM19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIM19 cDNA was amplified by PCR with the template pcxn2-GRIM19 and cloned into the shuttle plasmid pAdTrack-CMV. The plasmid pAdTrack-CMV-GRIM19 was linearized by PmeⅠ and homologously recombined with bone plasmid pAdEasy-1 in BJ5183, followed by identification by enzyme digestion. After transfection of linearized pAd-GRIM19 with PacⅠ into HEK293 cells, Ad-GRIM19 was obtained and amplified by 3 circles. SW480 cells were infected with Ad-GRIM19. The apoptosis rate was detected by flow cytometry. Agarose electrophoresis revealed the bands of recombinant plasmids identified by enzyme digestion were in the right range corresponding with expectation. Under the fluorescent microscopy, the package of Ad-GRIM19 in HEK293 cells and the expression of Ad-GRIM19 in SW480 cells were observed. The transfection of Ad-GRIM19 into SW480 cells increased the apoptosis rate of SW480 cells as compared with controls, It was concluded that Ad-GRIM19 was successfully constructed and the overexpression of GRIM19 in colon cancer cell lines could promote the apoptotic cell death.
文摘Objective： To investigate the effect of DL-lysine acetylsalicylate on proliferation of colon carcinoma cells line sw480. Methods： After treatment of DL-lysine acetylsalicylate, the study was performed by observing sw480 colorectal cancer cells with phase contrast microscope, making growth curve, and examining the inhibition rate of sw480 cells with MTT assay. Results： The morphology of sw480 cells showed characteristics of apoptosis, the cell growth curve showed inhibited proliferation of sw480 cells when treated with DL-lysine acetylsalicylate （P〈0.05）. The rate of inhibition was upward when the drug concentration increased. Conclusion： DL-lysine acetylsalicylate for injection can inhibit the growth of sw480 colorectal cancer cells obviously in a dose dependent manner.
基金Supported by the Multigrain Production and Processing Characteristic Discipline Construction ProjectPostdoctoral Scientific Research Foundation of Heilongjiang Province of China(LBH-Q13132)
文摘[Objectives] The aim was to elucidate the pro-apoptosis mechanism of naphthoquinone derivative 2-octyl sulfoxide-1,4-naphthoquinone(OSNQ) on human colon cancer SW480 cells.[Methods]The cytotoxic effect of OSNQ on colon cancer SW480 cells was detected by MTT colorimetry.The pro-apoptotic effect of OSNQ on human colon cancer SW480 cells was detected by Annexin V-FITC/PI double staining.The changes in expression of apoptosis-related proteins were detected by Western blot.[Results]The results of MTT assay showed that OSNQ had a significant cytotoxic effect on colon cancer SW480 cells.The results of Western blot showed that OSNQ induced the apoptosis in colon cancer SW480 cells through promoting the expression of pro-apoptotic caspase-3 and inhibiting the expression of apoptosis-inhibiting protein Bcl-2.[Conclusions] OSNQ has a significant cytotoxic effect on colon cancer SW480 cells,and it induces the apoptosis of colon cancer SW480 cells by AKT signaling pathway.
文摘目的观察青藤碱对高侵袭性人结肠癌细胞株SW 480移植瘤细胞周期及结肠瘤体组织中环氧合酶-2(COX-2)表达的影响,研究青藤碱防治结肠癌的机制。方法先用5只免疫功能正常的BALB/c-nu/nu裸小鼠建立高侵袭性人结肠癌瘤源,3周后取直径约2 mm的瘤块移植于20只免疫缺陷的BALB/c-nu/nu2裸小鼠回盲浆膜凹龛部,1周后再随机分为结肠癌组和青藤碱组。青藤碱组按10 m L/(kg·d)灌10%青藤碱治疗30 d,结肠癌组灌0.9%氯化钠注射液对照。测量BALB/c-nu/nu裸小鼠结肠瘤体积、瘤质量并计算肿瘤抑制率;流式细胞仪检测结肠瘤体组织SW 480细胞在G1、G2、M和S期的细胞周期比例;免疫组化法检测结肠瘤体组织COX-2的表达,RT-PCR法检测COX-2 m RNA表达。结果与结肠癌组比较,青藤碱组结肠瘤体积、瘤质量及S期细胞比值明显降低、G1、G2期细胞比值提高,肿瘤抑制率为(39.75%)(P<0.05);COX-2和COX-2 m RNA低表达(P<0.05)。结论青藤碱可能在G1、G2和S期诱导SW 480细胞阻滞、抑制结肠癌细胞株SW 480细胞的有丝分裂,并通过下调COX-2表达而间接影响癌细胞的增殖,具有抗高侵袭性人结肠癌的作用。
