Inhibitory effect of royal jelly acid on proliferation of human colon cancer SW620 cells and its network pharmacological analysis / 吉林大学学报(医学版)

Objective:To discuss the effect of royal jelly acid(10-HDA)on the proliferation and migration of the human colon cancer SW620 cells based on the network pharmacology,and to clarify its related molecular mechanism.Methods:The active ingredients such as 10-HDA and their corresponding targets were retrieved by using the keyword"royal jelly"from the Traditiomal Chinese Medicine Systems Pharmacology(TMSCP)Database and the Traditiomal Chinese Medicine Integrated Database(TCMID);the small molecule targets were predicted by the Swiss Target Prediction Database.The GeneCards Database and the Online Mendelian Inheritance in Man(OMIM)Database were used to obtain the targets with the keyword"Colon Cancer";the protein-protein interaction(PPI)network was constructed by using the String Database and Cytoscape 3.8.0 Software to screen the core targets;the Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were analyzed by Metascape Database;the specific ingredient 10-HDA was screened for the in vitro activity experiments.The human colon cancer SW620 cells with good growth status were divided into control group and different doses(1,5,10,15,and 20 mmol·L-1)of 10-HDA groups.The viabilities of the cells in various groups were detected by MTT method and the survival rates of the cells were calculated.The SW620 cells were divided into control group,low dose(5 mmol·L-1)of 10-HDA group,middle dose(10 mmol·L-1)of 10-HDA group,and high dose(15 mmol·L-1)of 10-HDA group;Hoechst33342 staining method was used to observe the morphology of the cells in various groups;cell scratch test was used to detect the scratch healing rates of the cells in various groups;flow cytometry was used to detect the percentages of the cells at different cell cycles in various groups;biochemical method was used to detect the activities of total antioxidant capacity(T-AOC)and superoxide dismutase(SOD)in the cells in various groups;Western blotting method was used to detect the expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cysteine-containing aspartate proteolytic enzyme-3(Caspase-3),cysteine-containing aspartate proteolytic enzyme-9(Caspase-9),glycogen synthase kinase 3β(GSK3β),β-catenin,and cyclin D1 proteins in the cells in various groups.Results:Six active ingredients of royal jelly were screened out by the TCMSP Database,and 28 core targets of 10-HDA in the treatment of colon cancer were obtained.The GO function enrichment analysis mainly included the signaling pathways such as cell proliferation and apoptosis.The KEGG signaling pathway enrichment analysis included the cell cycle,prostate cancer,cell senescence,and p53 signaling pathways;the GSK3β/β-catenin signaling pathway was closely related to the cell cycle.Compared with control group,the viabilities of the cells in 5,10,15,and 20 mmol·L-110-HDA groups were decreased in a dose-dependent manner(P<0.05 or P<0.01),the numbers of apoptotic cells in different doses of 10-HDA groups were significantly increased,and the scratch healing rates of the cells were significantly decreased(P<0.05 or P<0.01);the percentages of the cells at S phase in middle and high doses of 10-HDA groups were significantly increased(P<0.05 or P<0.01),the activities of T-AOC and SOD in the cells in different doses of 10-HDA groups were significantly decreased(P<0.05 or P<0.01).Compared with control group,the expression level of Bcl-2 protein in the cells in low dose of 10-HDA group was significantly decreased(P<0.01),and the expression level of GSK3β protein was significantly increased(P<0.05);compared with control group,the expression levels of Bax,Caspase-3,Caspase-9,and GSK3β proteins in the cells in middle and high doses of 10-HDA groups were significantly increased(P<0.05 or P<0.01),and the expression levels of Bcl-2,β-catenin,and CyclinD1 proteins were significantly decreased(P<0.01).Conclusion:10-HDA can significantly inhibit the proliferation and migration of the colon cancer cells and promote the apoptosis and oxidation levels of the colon cancer cells,and its mechanism may be related to the activation of the GSK3β/β-catenin signaling pathway.

Expression of TRAP1 in colon cancer tissues and its relationship with clinicopathological features and patient prognosis and the possible molecular mechanisms / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨肿瘤坏死因子受体相关蛋白1（TRAP1）在结肠癌组织和细胞中的表达及其与临床病理特征和患者预后的关系和相关分子机制。方法：通过TCGA和GEO数据全面分析TRAP1在结肠癌中的表达及其与临床病理特征和患者预后的关系，选取2020年10月至2021年03月间在山西医科大学第一医院手术切除的10例结肠癌组织及相应癌旁组织标本，用IHC染色法检测中国人结肠癌组织中TRAP1的表达进行验证，运行R包（survival和survminer）进行Kaplan-Meier生存分析；在线分析TRAP1蛋白的信号肽及穿膜结构域，通过基因富集分析软件进行GO分析和KEGG分析。培养结肠癌SW480和SW620细胞，将si-NC和si-TRAP1转染结肠癌细胞，实验分为空白对照组、si-NC组和si-TRAP1组，采用qPCR法检测转染后各组结肠癌细胞中TRAP1的表达，FCM检测转染后各组细胞的细胞周期和凋亡情况。结果：与癌旁组织比较，TRAP1在结肠癌组织中呈高表达（P<0.01），TRAP1表达水平与淋巴结转移有关联（P<0.05），TRAP1高表达组结肠癌患者5年OS率较低（P<0.05）。TRAP1蛋白属于细胞质蛋白，功能富集结果显示TRAP1及其相关分子与细胞周期、核糖体生物发生等信号通路有关（均P<0.01），TRAP1高表达组的结肠癌代谢重编程基因簇和线粒体蛋白输入基因簇水平升高（均P<0.01）。敲减TRAP1后，结肠癌细胞周期阻滞于G1期，细胞凋亡水平显著升高（均P<0.01）。结论：TRAP1在结肠癌组织中呈高表达，且与患者淋巴结转移和低OS率相关联，敲减TRAP1可阻滞结肠癌细胞周期并促进其凋亡。

Alterations in APC, BECN1, and TP53 gene expression levels in colon cancer cells caused by monosodium glutamate / Alterações nos níveis de expressão gênica APC, BECN1 e TP53 em células de câncer de cólon causadas por glutamato monossódico

Colorectal cancer (CRC) is a disease with high incidence worldwide. As of 2018, it is the second leading cause of cancer deaths in the world. In Saudi Arabia, the incidence of this disease has been increasing in the younger population. Both genetic and lifestyle factors may have contributed to its increased incidence and pathogenesis. Monosodium glutamate (MSG) is a food flavor enhancer that can be found in many commercial foods, and it can sometimes be used as a substitute to table salt. MSG has been investigated for its possible genotoxicity, yielding controversial results. In the present study, the effect of MSG on cell viability and its effect on expression of APC, BECN1, and TP53 genes in SW620 and SW480 colon cancer cell lines were studied. TP53 is a tumor suppressor gene that functions in modifying DNA errors and/or inducing apoptosis of damaged cells, and both APC and BECN1 genes are involved in CRC and are of importance in cellular growth and metastasis. Cancer cell viability was analyzed using MTT assay, and the results showed a significant increase in the number of viable cells after 24h of treatment with MSG with different concentrations (0.5, 1.0, 10, 50, and 100mM). Moreover, gene expression results showed a significant increase in the expression levels of APC and BECN1 under specified conditions in both cell lines; conversely, TP53 showed a significant decrease in expression in SW620 cells. Thus, it can be concluded that MSG possibly confers a pro-proliferative effect on CRC cells.

O câncer colorretal (CCR) é uma doença com alta incidência mundial. Desde 2018, é a segunda principal causa de mortes por câncer no mundo. Na Arábia Saudita, a incidência dessa doença vem aumentando na população mais jovem. Tanto fatores genéticos quanto de estilo de vida podem ter contribuído para o aumento da sua incidência e patogênese. O glutamato monossódico (MSG) é um intensificador de sabor de alimentos que pode ser encontrado em muitos alimentos comerciais e às vezes pode ser usado como um substituto do sal de cozinha. O MSG tem sido investigado por sua possível genotoxicidade, produzindo resultados controversos. Neste estudo, foram estudados o efeito do MSG na viabilidade celular e seu efeito na expressão dos genes APC, BECN1 e TP53 em linhas de células de câncer de cólon SW620 e SW480. TP53 é um gene supressor de tumor que atua modificando erros de DNA e/ou induzindo apoptose de células danificadas, estando os genes APC e BECN1 envolvidos no CRC e sendo importantes no crescimento celular e metástase. A viabilidade das células cancerosas foi analisada por meio do ensaio MTT, e os resultados mostraram um aumento significativo no número de células viáveis após 24 h de tratamento com MSG em diferentes concentrações (0,5; 1,0; 10; 50 e 100mM). Além disso, os resultados da expressão gênica mostraram um aumento significativo nos níveis de expressão de APC e BECN1 sob condições especificadas em ambas as linhagens celulares. Por outro lado, TP53 mostrou uma diminuição significativa na expressão em células SW620. Assim, pode-se concluir que, possivelmente, o MSG confere um efeito pró-proliferativo às células CRC.

Alterations in APC, BECN1, and TP53 gene expression levels in colon cancer cells caused by monosodium glutamate

Abstract Colorectal cancer (CRC) is a disease with high incidence worldwide. As of 2018, it is the second leading cause of cancer deaths in the world. In Saudi Arabia, the incidence of this disease has been increasing in the younger population. Both genetic and lifestyle factors may have contributed to its increased incidence and pathogenesis. Monosodium glutamate (MSG) is a food flavor enhancer that can be found in many commercial foods, and it can sometimes be used as a substitute to table salt. MSG has been investigated for its possible genotoxicity, yielding controversial results. In the present study, the effect of MSG on cell viability and its effect on expression of APC, BECN1, and TP53 genes in SW620 and SW480 colon cancer cell lines were studied. TP53 is a tumor suppressor gene that functions in modifying DNA errors and/or inducing apoptosis of damaged cells, and both APC and BECN1 genes are involved in CRC and are of importance in cellular growth and metastasis. Cancer cell viability was analyzed using MTT assay, and the results showed a significant increase in the number of viable cells after 24 h of treatment with MSG with different concentrations (0.5, 1.0, 10, 50, and 100mM). Moreover, gene expression results showed a significant increase in the expression levels of APC and BECN1 under specified conditions in both cell lines; conversely, TP53 showed a significant decrease in expression in SW620 cells. Thus, it can be concluded that MSG possibly confers a pro-proliferative effect on CRC cells.

Resumo O câncer colorretal (CCR) é uma doença com alta incidência mundial. Desde 2018, é a segunda principal causa de mortes por câncer no mundo. Na Arábia Saudita, a incidência dessa doença vem aumentando na população mais jovem. Tanto fatores genéticos quanto de estilo de vida podem ter contribuído para o aumento da sua incidência e patogênese. O glutamato monossódico (MSG) é um intensificador de sabor de alimentos que pode ser encontrado em muitos alimentos comerciais e às vezes pode ser usado como um substituto do sal de cozinha. O MSG tem sido investigado por sua possível genotoxicidade, produzindo resultados controversos. Neste estudo, foram estudados o efeito do MSG na viabilidade celular e seu efeito na expressão dos genes APC, BECN1 e TP53 em linhas de células de câncer de cólon SW620 e SW480. TP53 é um gene supressor de tumor que atua modificando erros de DNA e/ou induzindo apoptose de células danificadas, estando os genes APC e BECN1 envolvidos no CRC e sendo importantes no crescimento celular e metástase. A viabilidade das células cancerosas foi analisada por meio do ensaio MTT, e os resultados mostraram um aumento significativo no número de células viáveis após 24 h de tratamento com MSG em diferentes concentrações (0,5; 1,0; 10; 50 e 100mM). Além disso, os resultados da expressão gênica mostraram um aumento significativo nos níveis de expressão de APC e BECN1 sob condições especificadas em ambas as linhagens celulares. Por outro lado, TP53 mostrou uma diminuição significativa na expressão em células SW620. Assim, pode-se concluir que, possivelmente, o MSG confere um efeito pró-proliferativo às células CRC.

Betulinic Acid Induces Autophagy and Apoptosis of Human Colorectal Cancer SW620 Cells by Regulating PI3K/Akt/mTOR Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of betulinic acid （BA） on apoptosis and autophagy of human colorectal cancer SW620 cells and the regulatory role of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodCell viability was detected by methyl thiazolyl tetrazolium （MTT） colorimetry to determine the optimal administration time and dosage for subsequent experiments. Four groups were designed， including blank group and low-， medium-， and high-dose BA groups. Hematoxylin-eosin （HE） staining was conducted for the observation of SW620 cell morphology， and annexin-V/propidium iodide double staining for the determination of apoptosis rate in SW620 cells. Hoechst33258 staining and MDC staining were used for the observation of apoptosis and autophagy， respectively. Western blotting was employed to determine the protein levels of B-cell lymphoma/leukemia-2（Bcl-2）-associated X protein （Bax）， aspartate proteolytic enzyme-9 （Caspase-9）， activated aspartate proteolytic enzyme-3 （cleaved Caspase-3）， microtubule-associated protein 1 light chain 3 （LC3）， the mammalian homolog of yeast Atg6 （Beclin-1）， p62， phosphorylated PI3K （p-PI3K）， phosphorylated Akt （p-Akt）， and phosphorylated mTOR （p-mTOR） in SW620 cells. ResultBA inhibited the activity of SW620， HT29， and HCT116 cells in a concentration- and time-dependent manner. The cells treated with BA for 48 h had lower viability than those treated for 24 h （P<0.05， P<0.01）. The half maximal inhibitory concentration (IC50） value of BA at the time point of 48 h was also lower than that at the time point of 24 h （P<0.01）， and that for SW620 cells was the minimum. BA induced the apoptosis in a concentration-dependent manner and increased the autophagosomes. Compared with the blank group， BA increased the apoptosis rate （P<0.01）， up-regulated the protein levels of Bax， Caspase-9， cleaved Caspase-3， and LC3 Ⅱ （P<0.05， P<0.01）， and down-regulated the protein levels of p62， p-Akt， p-PI3K， and p-mTOR （P<0.01）. Additionally， medium- and high-dose BA up-regulated the protein level of beclin-1 （P<0.01）. ConclusionBA may inhibit the activity of SW620 cells by hindering the PI3K/Akt/mTOR signaling pathway to induce cell apoptosis and autophagy.

lncRNA SNHG10 promotes the invasion and migration of colorectal cancer SW620 cells by targeting miR-532-3p / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of long non-coding RNA (lncRNA) small nucleolar RNA host gene 10 (SNHG10) in colorectal cancer tissues and cells and its effect on the invasion and migration of colorectal cancer cells and the underlying mechanism. Methods: From January 2018 to December 2019, 78 pairs of colorectal cancer tissue and para-cancerous tissues from the patients who had radical colorectal cancer resection in Henan Provincial People's Hospital were collected. Quantitative PCR (qPCR) was performed to quantify the levels of lncRNA SNHG10 and miR-532-3p in colorectal cancer tissues, colorectal cancer cell lines (SW480, SW620, HT-29 and LoVo) and human normal colorectal mucosal FHC cells; and their correlations with the clinicopathological features of colorectal cancer patients were further analyzed. Dual-luciferase reporter gene assay was used to validate the targeted relationship between lncRNA SNHG10 and miR-532-3p. After transfection with si-SNHG10 or miR-532-3p mimic or co-transfection of si-SNHG10 and miR-532-3p inhibitor, the invasion and migration of SW620 cells were detected by Transwell assay, and the protein expression of E-cadherin, N-cadherin and vimentin were detected by WB. Results: SNHG10 was highly expressed in colorectal cancer tissues and cells (all P<0.05), and its expression was related to TNM stage and distant metastasis (all P<0.05). miR-532-3p was lowly expressed in colorectal cancer tissues and cells, and its expression was correlated with TNM stage, lymphonode metastasis and distant metastasis (all P<0.05). The expression of SNHG10 and miR-532-3p in colorectal cancer tissues was negatively correlated (r=-0.225, P=0.048). Dual-luciferase reporter gene assay confirmed that SNHG10 targetedly regulated miR-532-3p. Both down-regulation of SNHG10 and up-regulation of miR-532-3p significantly inhibited the invasion and migration of SW620 cells (all P<0.05), up-regulated the expression of E-cadherin (P<0.05), while down-regulated the expression of N-cadherin and vimentin (all P<0.05). After transfection with miR-532-3p inhibitor, the inhibitory effect of knocking down the expression of lncRNA SNHG10 on the invasion and migration of colorectal cancer cells was reversed (all P<0.05). Conclusions: LncRNA SNHG10 is highly expressed in colorectal cancer and is associated with TNM stage and distant metastasis. LncRNA SNHG10 affects the invasion and metastasis of colorectal cancer cells by targeting miR-532-3p and regulating EMT.

Effect of Schisandrin B on Proliferation and Migration of Human SW620 Colon Cancer Cell Via VEGF/PI3K/Akt Signaling Pathway / 中国药学杂志

OBJECTIVE: To study the changes of VEGF signaling pathway in colon cancer patients and the effect of schisandrin B on SW620 cells and to analyze its possible mechanism. METHODS: The protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt in human cancerous colon samples and adjacent normal samples were detected by Western blotting. The proliferation of SW620 cells was detected by CCK-8 method. The mRNA expression of VEGFA, VEGF-R2, PI3K and Akt in SW620 cells were detected by real-time PCR. The protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt in SW620 cells were detected by Western blotting. RESULTS: The protein expression of VEGF-R2, PI3K, Akt and p-Akt in human cancerous colon samples was significantly higher than that in the adjacent normal samples(P<0.05). Compared with the control group, schisandrin B could significantly inhibit the proliferation and migration of SW620 cells, the mRNA expression of VEGFA, VEGF-R2, PI3K and Akt(P<0.01) and the protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt(P<0.05) in SW620 cells also were significantly decreased by schisandrin B. CONCLUSION: The VEGF/PI3K/Akt signaling pathway is activated in colon cancer patients. Schisandrin B could inhibit the activity and migration of SW620 cells and inhibit the VEGF/PI3K/Akt signaling pathway.

Id1 and Id3 synergistically influence invasion and migration of colon cancer SW620 cells by inducing EMT / 中国肿瘤生物治疗杂志

@# Objective: To investigate whether inhibitor of differentiation 1 gene (Id1) and Id3 gene can synergistically promote epithelial-mesenchymal transition (EMT), invasion and migration of colon cancer SW620 cells and to explore its underlying mechanisms. Methods: The SW620 cell strain with Idl or Id3 gene knockdown and the SW620 cell strain with Id1/Id3 gene double-knockdown were constructed by lentiviral vectors transfection. The SW620 cells were divided into four groups, which included SW620-Sh-Id1 group (transfected with shRNA-Id1), SW620-Sh-Id3 group (transfected with shRNA-Id3), SW620-Sh-Id1-Id3 group (transfected with shRNAId1 plus shRNA-Id3) and SW620-NC group (transfected with negative lentivirus). The efficiency of knockdown was detected by Realtime qPCR and Western blotting. The influence of stable knockdown of Idl or Id3 on cell morphological change was observed under a microscope. The changes of migration and invasion abilities of the SW620 cells were determined by wound healing assay and Transwell assay. EMT, invasion and migration related proteins were measured by Western blotting. Results: The SW620 cell strains with Idl and/or Id3 gene knockdown were successfully constructed. Idl and Id3 knockdown induced the epithelial-like to the mesenchymanl-like transformation of SW620 cells. (1) Compared with the control group, the invasion and migration abilities of the SW620 cells were significantly decreased in the SW620-Sh-Id1 group and SW620-Sh-Id3 group (all P<0.05). (2) Meanwhile, the invasion and migration abilities in the SW620-Sh-Id1-Id3 group were obviously weaker than the SW620-Sh-Id1 group and SW620-Sh-Id3 group (all P<0.05). (3) Compared with the control group, the SW620-Sh-Id1 group and SW620-Sh-Id3 group had a reduction in the protein expressions of βcatenin, snail1 and MMP2, and an increase in the protein expressions of E-cadherin and TIMP2 (all P<0.05). (4) Compared with the SW620-Sh-Id1 group and SW620-Sh-Id3 group , the protein expressions of β-catenin, snail1 and MMP2 were reduced, and the protein expressions of E-cadherin and TIMP2 were increased in the SW620-Sh-Id1-Id3 group (all P<0.05). Conclusion: Id1 and Id3 could synergistically influence invasion and migration of SW620 cells, possibly through inducing EMT.

Differential analysis of gene expression profiles for lymphonode metastasis of colon cancer / 临床检验杂志

Objective To investigate the differences in the gene expression profiles between SW480 and SW620 cell lines.Methods A dataset of GDS756 containing the gene expression profiles of SW480 and SW620 was downloaded from the GEO database in NCBI.The differential expression genes between SW480 and SW620 were analyzed with gene set enrichment analysis (GSEA) and leading edge subset analysis.The genes in leading edge subset were re-annotated by FunRich software.The core genes of leading edge subset closely relating to SW480 or SW620 were analyzed with the STRING on-line analytical system.The functional core genes closely relating to SW480 or SW620 were obtained by the combined analysis of the core genes and high frequency genes from leading edge subset.Results GSEA identified 12 significantly enriched gene sets,491 leading edge genes and 7 highly overlapping genes from SW480 and 80 significantly enriched gene sets,870 leading edge genes and 6 highly overlapping genes from SW620.The STRING system identified 5 core genes from SW480 and 8 from SW620.The combined analysis of GSEA and bionetwork obtained 2 functional core genes,TOP2A and CDK1,from SW620.Conclusion The SW480 and SW620 cells with identical genetic background have different functional gene expression profiles,and the functional core genes TOP2A and CDK1 in SW620 cells may be related to the signal pathways of colon cancer metastasis.

Effect of oxymatrine on p16/cyclinD1/CDK4 pathway in human colon adenocarcinoma SW620 cells / 中草药

Objective: To explore the antitumor effect of oxymatrine and detect the mechanism involved. Methods: The Anti-proliferative effects of oxymatrine in human colon adenocarcinoma SW620 cells were assessed using MTT assay. SW620 cells treated with oxymatrine were assessed with Hoechst 33258 staining and cell cycle distribution assay was performed by flow cytometry. The quantitative real-time PCR assay was used to evaluate the expression of p16, cell cycle-related cyclinD1 and CDK4 mRNA at the genetic level. To investigate the molecular mechanisms underlying alterations in cell apoptosis, the proteins p16, cell cycle-related cyclinD1, and CDK4 were determined by Western blotting analysis. Results: Oxymatrine could significantly inhibit the growth of SW620 cells compared with the control group, the IC50 was 4.02 μmol/L. Its anticancer activity was related to the alteration in expression of p16, cyclinD1, and CDK4 (P<0.05, 0.01). Conclusion: These results suggest that oxymatrine produces the obvious antitumor effects on SW620 cells in vitro, induces the cell arrest in G1 phase which is related to the regulation on the protein expression of p16, cyclinD1, and CDK4.

Effects of miR-192 on the expression of ZEB2 and the abilities of migration and invasion of colorectal cancer cells / 肿瘤

Objective: To investigate the effects of miR-192 on the expression of zinc-finger E-box binding homeobox 2 (ZEB2) and the abilities of migration and invasion of colorectal cancer (CRC) SW480 and SW620 cells. Methods: After transfection of miR-192 mimics into SW480 and SW620 cells, the expression levels of miR-192 and ZEB2 mRNA and ZEB2 protein were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The abilities of migration and invasion of SW480 and SW620 cells after transfection with miR-192 mimics were determined by wound healing and Transwell assays, respectively. The recombinant vector pmiR-ZEB2-wt and miR-192 mimics were co-transfected into SW480 cells. The binding site of 3′-untranslated region (3′-UTR) of ZEB2 gene with miR-192 was verified by Dual Luciferase™ Reporter Gene Assay. Results: As compared with the negative control (NC) group (transfection with miR-192 mimics-NC), the expression levels of miR-192 in SW480 and SW620 cells after transfection with miR-192 mimics were increased (P < 0.05), and the expression levels of ZEB2 mRNA and protein were decreased (P < 0.05), as well as the abilities of migration and invasion of SW480 and SW620 cells were inhibited (P < 0.05). The Dual Luciferase™ Reporter Gene Assay revealed that the luciferase activity in SW480 cells after co-transfection with recombinant vector pmiR-ZEB2-wt and miR-192 mimics was inhibited (P < 0.05), which indicated that the 3′-UTR of ZEB2 harbored a binding site for miR-192. Conclusion: ZEB2 may be one of the target genes for miR-192. Overexpression of miR-192 may down-regulate the ZEB2 expression and inhibit the migration and invasion of SW480 and SW620 cells. Copyright© 2014 by TUMOR.

Inhibitory effect of platycodin D on human colonic cancer SW620 cell proliferation / 中国药学杂志

OBJECTIVE: To explore the inhibitory effect and its mechanism of platycodin D (PD) on human colonic cancer SW620 cell proliferation. METHODS: The inhibitory effect of PD on SW620 cell proliferation was analyzed by MTT assay, while the effect of PD on cell cycle distribution and apoptosis were evaluated by flow cytometry. The effect of PD on expression of cyclin and ap-optosis associated proteins were detected by Western blot. RESULTS: The cell proliferation of human colonic cancer SW620 cell was inhibited by PD in a dose-dependent manner. After cells were treated with PD(15, 20 μmol · L-1) for 24 h, the proportions of cells in G0/G1 phase were (72.83±5.26)% and (76.82±5.83)% respectively, while the control group cells was (56.78±4.92)%, which suggested PD could block SW620 in the G, phase compared with the control group cells. Furthermore, the expressions of cyclinD1, c-myc and GDK6 were reduced obviously. Compared with the control group cells, the apoptotic rates were increased [(19.5±5.1)%, (35.8±5.3)% and (43.8±4.0)% respectively] after cells were treated by PD (10, 15, 20 μmol · L-1) for 48 h. The expression of procaspase 3 and PARP with proenzyme form were reduced. CONCLUSION: PD could inhibit the growth of colon cancer cell by blocking SW620 in the G, phase through regulating the expression of cyclinD1, c-myc and CDK6 and thus inducing the apoptosis by cell cycle arrest.

PLCE1 over-expression inhibits migration of colon cancer SW620 cells and induces their apoptosis / 肿瘤

Objective: To investigate the effects of phospholipase C epsilon-1 (PLCE1 ) over-expression on migration, cell cycle and apoptosis of human colon cancer cells. Methods: The SW620 cells overexpressing PLCE1 were constructed through lipofection. Three groups were designed as follows: parent group (without transfection), control group (transfected with empty plasmid containing green fluorescent protein) and experimental group (transfected with pcDNA-DEST53-PLCE1 plasmid). The expression levels of PLCE1 mRNA and protein in SW620 cells were detected by real-time fluorogenic quantitative-PCR (RFQ-PCR) and Western blotting, respectively. The effect of PLCE 1 over-expression on migation ability of SW620 cells was detected by Transwell chamber assay. The cell cycle distribution and apoptosis rate of SW620 cells were detected by flow cytometry (FCM). The apoptosis was analyzed by using DNA ladder method. Results: The migration ability of SW620 colon cancer cells was inhibited by over-expression of PLCE 1. The numbers of migrated cells in the parent, control and experimental groups were 32.60±2.42, 32.20±3.25 and 8.80±1.72, respectively, and the difference among three groups was significant (P < 0.01). The over-expression of PLCE1 prolonged phase G1 and induced apoptosis. Conclusion: PLCE1 over-expression can inhibit the migration ability of colon cancer cells and induce their apoptosis. PLCE1 over-expression can reduce the malignant degree of colon cancer cells, and this gene may be a new antioncogene related to colon cancer. Copyright© 2011 by TUMOR.

The effect of silencing Sp1 gene on proliferation of colon carcinoma cell SW620 by RNA interference / 肿瘤研究与临床

Objective To inhibit the expression of transcription factor special protein 1(Sp1) through RNA interference (RNAi) technique and to investigate its impact on the proliferation ability of colorectal cancer cell line SW620. Methods The recombinant plasmid of Sp1 RNAi (pGenesil-1-Sp1) was constructed and transfected into SW620 cells by Lipofectamine. The transfcction efficiency was observed under fluorescence confocal microscopy. Expression levels of Sp1 mRNA and protein from SW620 after transfection were examined by real time PCR and Western blot respectively, after transduction of the recombinant plasmid into the SW620. The proliferation ability of SW620 cell line was evaluated by MTT assay. Results The expression plasmid (pGenesil-1-Sp1) against Sp1 was successfully constructed, recombinant vectors could reduce the expressions of Sp1 mRNA and protein in SW620, the ratio of inhibition of the expression of Sp1 mRNA and protein was 68.47 % and 73.82 % in 48th hour respectively. Compared with the control group, the difference was significant (P <0.05). MTT showed that the proliferation ability of SW620 cell was degraded. Conclusion Silencing Sp1 gene by the RNAi technology can actively inhibit the proliferation of SW620 cell. The successful application of Spl SiRNA extends the list of available therapeutic modalitics in the treatment of human colon cancer.

Inhibitory effects of AP-2α on proliferation of colon cancer cell of SW620 and its mechanism / 肿瘤研究与临床

Objective To study inhibitory effects of transcription factor activator protein-2α(AP-2α)on proliferation of colon cancer cells in vitro and its mechanism. Methods The peDNA3.1 (+)-AP-2α recombinant plasmid was constructed. Plasmid pcDNA3.1(+)- AP-2α and pcDNA3.1(+)was transfected into SW620 cell by liposome mediation for transient expression, and proliferative activities of SW620 cell were evaluated by MTT assay. The change in the mRNA and protein expression level of ER-β before and after transfection was detected using the methods of Real-Time PCR and Western blotting respectively. Results The mRNA and protein expressions of AP-2α could be enhanced by transfecting of AP-2α gene in SW620 cell. MTT assay indicated: the proliferation velocity of SW620 cell for transfection of the pcDNA3.1(+)-AP-2α plasmid was apparently inhibited. The expression of ER-β in SW620 cell increased significantly after AP-2α gene transfection. Compared with control group, the difference was significant (P＜0.05). Conclusion Overexpression of AP-2α inhibits the proliferation of SW620 cell in vitro, which is probably related with activation of ER-β.

Effects of AP-2? on invasive growth and ER-? expression of colon cancer cell line / 中国肿瘤生物治疗杂志

Objective: To study the effects of transcription factor activator protein-2?(AP-2?)on invasive growth and estrogen receptor-?(ER-?) expression in human colon cancer SW620 cells,and to probe into the involved molecular mechanism.Methods: Plasmid pcDNA3.1(+)-AP-2? and pcDNA3.1(+) were transfected into SW620 cells by liposome-mediated transfection.The adhesion,invasion and migration abilities of SW620 cells were measured by metrical gel adhesion assay and modified Boyden chamber(Transwell assay).The gene and protein expression levels of AP-2? and ER-? in SW620 cells were examined by Real-time PCR,Western blotting and immunofluorescence cytochemistry.The interaction between AP-2? DNA and ER-? in SW620 cells was measured by electrophoretic mobility shift assay(EMSA) after AP-2? gene transfection.Results: Overexpression of AP-2? markedly reduced the adhesion,invasion and migration abilities of SW620 cells(all P

Effect of sophoridine on proliferation and apoptosis of human colon adenocarcinoma cells(SW620) / 中国药理学通报

Aim To investigate the growth-inhibition and the mechanism of apoptosis induced by sophoridine(SRI) on SW620 cells.Method MTT assay was used to detect the half-inhibition concentration(IC50),and fluorescence microscopy,electron microscopy,DNA fragmentation analysis,flow cytometry(FCM) were used to demonstrate the presence of apoptosis.Results SRI inhibited the growth of SW620 cells significantly in a dose-and time-dependent manner,and morphological characteristics of apoptosis were observed with condensation of nucleus,bubble of cytoplasm,fragment of nucleus.A DNA ladder pattern of internucleosomalfragmentation was observed.Compared with that of control group,the percentage of the G0/G1 phase and the Sphase cells increased after treated by SRI.SW620 cells were induced to undergo apoptosis and underwent G0/G1 arrest with exposure to SRI shown by FCM.Conclusions Sophoridine could induce the inhibition of cell growth by means of apoptosis in a dose-and time-dependent manner,and the blocking of G0/G1 phase of cells was involved in the mechanism of apoptosis.