Effects and Mechanism of LASP1 on Proliferation, Migration, and Apoptosis of Human Colorectal Cancer LOVO Cells / 肿瘤防治研究

Objective To explore the effects and mechanism of LASP1 gene expression on the proliferation, migration, and invasion of human colorectal cancer (LOVO) cells. Methods LASP1 overexpression plasmids and LASP1 interference plasmids were constructed and transfected to LOVO cells. qRT-PCR was used to detect LASP1 mRNA expression and validate the transfection. MTT method and Tunel staining were used to detect cell proliferation and apoptosis, respectively, and scratch test and Transwell test were employed to determine the migration and invasion abilities of cells. Western blot was applied to analyze the expression of LASP1, p-FAK/FAK, and p-AKT/AKT protein in cells. Results The plasmids were successfully transfected. LASP1 overexpression increased the proliferation, migration, and invasion of LOVO cells, decreased the apoptosis, and increased LASP1, p-FAK/FAK, p-AKT/AKT protein expression (P < 0.01). LASP1 knockdown reduced the proliferation, migration, and invasion of LOVO cells, increased the apoptosis, and decreased LASP1, p-FAK/FAK, and p-AKT/AKT protein expression (P < 0.01). Conclusion LASP1 positively regulates the FAK/AKT signaling pathway to promote the proliferation, migration, and invasion of LOVO cells.

Glia maturation factor-gamma inhibits the proliferation of human colorectal cancer LoVo cells and affects the cytoskeletal motion of human umbilical vein endothelial cells / 中国组织工程研究

BACKGROUND: Glial maturation factor-gamma (GMFG) is one of the members of the ADF/cofilin superfamily protein in the process of cytoskeleton remodeling. It can regulate the cell movement by regulating actin-mediated cytoskeleton reorganization. The authors previously found that the high expression of GMFG is associated with the clinicopathological features of colorectal cancer patients, but the specific upstream and downstream mechanisms need to be explored. OBJECTIVE: To study the effects of GMFG on cytoskeleton and LoVo cell proliferation. METHODS: (1) Human umbilical vein endothelial cells were inoculated in a laser confocal dish. After the cells grew stably, the correlation between the expression of GMFG and the expression of cytoskeletal protein F-actin was observed by immunofluorescence staining. Colchicine inhibited mitosis. The cells were divided into control group, 0.5 mg/L colchicine group and 1.0 mg/L colchicine group. The cell cycle was observed by flow cytometry and GMFG protein expression was observed by western blot assay. (2) Human colorectal cancer LoVo cells were inoculated in a laser confocal dish. After the cells grew stably, the correlation between GMFG expression and cell cycle was observed by immunofluorescence staining. Afterwards, siRNA was used to interfere the expression of GMFG in LoVo cells. The LoVo cells were divided into blank control group, empty transfer group and siRNA-GMFG group. The cell proliferation rate was detected by Edu assay. RESULTS AND CONCLUSION: (1) In human umbilical vein endothelial cells, the expression of GMFG was associated with the cytoskeletal motion of human umbilical vein endothelial cells, and the expression of GMFG increased when the cytoskeleton retracted. After colchicine inhibited the mitosis of human umbilical vein endothelial cells, the proportion of cells in G2/M phase increased and the expression of GMFG protein decreased with the increase of colchicine dosage. (2) In LoVo cells, the expression intensity of GMFG was associated with mitosis, and the expression of GMFG was enhanced in the middle and late stages of cell mitosis. (3) After siRNA interference with GMFG in LoVo cells, the cell proliferation rate was decreased. (4) Results verify that interference with GMFG can inhibit the proliferation rate of LoVo cells of human colorectal cancer, and there is a certain correlation between cytoskeletal motion and GMFG expression.

The mechanism study of miRNA-19a impacting on biological characteristics of colon cancer LoVo cell / 国际检验医学杂志

Objective To investigate the mechanism of miRNA-19a impacting on biological characteristics of colon cancer Lovo cell.Methods miRNA-19a mimics was transfected into human colon cancer cell line LoVo cells by liposome mediated transfection.The expression of miRNA-19a gene after transfection was detected.Cell proliferation was detected by CCK8 Kit.The effect of apoptosis was detected by TUNEL method.In vitro invasiveness of cells was detected by cell invasion chamber method.Hsp90/Akt signaling pathways changes were detected by RT-PCR and Western blot.Results The expression of miRNA-19a were increased.The ability of proliferation were promoted in vitro.The cell apoptosis was reduced.The cell invasion was increased.The expression of Hsp90 and Akt gene and protein were increased.Conclusion miRNA-19a could enhance Lovo cell invasion ability by the method of raising the expression of Hsp90 and Akt signaling pathways,accelerating LoVo cell proliferation,reducing cell apoptosis.

Anti-proliferation effect of resveratrol and p38 MAPK in human colon cancer cells / 中国药理学通报

Aim To investigate the relationship be-tween the anti-proliferation effect of resveratrol ( Res ) and p38 MAPK in colon cancer cells .Methods Crys-tal violet staining , Western blot and flow cytometry were employed to analyze the effect of Res on the pro-liferation in LoVo cells.Western blot assay was used to detect the effect of Res on the apoptosis of LoVo cells and the phosphorylation of p 38 MAPK.Crystal violet staining and Western blot assay were used to analyze whether p38 MAPK was involved in the Res-induced proliferation inhibition and apoptosis in LoVo cells .Re-sults Res inhibited the proliferation , arrested cell cy-cle at S phase , and increased the protein level of PC-NA in LoVo cells apparently .Res increased the level of Bad in LoVo cells, but decreased the level of Bcl-2. Although Res exerted no substantial effects on total lev-el of p38 MAPK, it markedly increased the phospho-rylation level of p38 MAPK in LoVo cells.p38 MAPK inhibitor promoted the proliferation , and decreased the anti-proliferation effect of Res on LoVo cells .Moreo-ver , the effects of Res on the level of Bcl-2 and Bad were both reduced by the p 38 MAPK inhibitor .Con-clusions Res can inhibit the proliferation of LoVo cells, which may be partly mediated by promoting the phosphorylation of p38 MAPK.

Mechanism of ethyl gallate induced the apoptosis of Lovo cells through mitochodrial pathway / 中国药学杂志

OBJECTIVE: To study the effect of ethyl gallate in inducing apoptosis of human colon carcinoma Lovo cells. METHODS: Cultured human colon carcinoma Lovo cells in vitro were used as the research object. Morphological changes of apoptotic cells were observed by inverted microscope.AnnexinV-FITC/PI assay and DNA Ladder assay were performed to study the apoptotic effects. Immunocytochemistry was used to detect the expression of cleaved caspase-9 and cleaved caspase-3 proteins. RESULTS: Ethyl gallate had growth inhibition on colon cancer Lovo cells,the density was decrease: the apoptotic rates were increased with dose increase: Agarose gel electrophoresis showed evident DNA fragmentation: the expression of cleaved caspase-9 and cleaved caspase-3 proteins was increased in treated groups. CONCLUSION: Ethyl gallate could induce the apoptosis in Lovo cells through up-regulating the expression of cleaved caspase-9 and cleaved caspase-3 protein,and then activated the mitochondrial pathway.

Study on relationship between anti-proliferation effect of resveratrol and resveratrol-induced bone morphogenetic protein 9 and its receptors in colon cancer cells / 中国药理学通报

Aim To study the anti-proliferation effect of resveratrol (Res)and the role of Res-induced bone morphorgenetic protein 9 (BMP9 )in this process in colon cancer cells.Methods Crystal violet staining and flow cytomtry were introduced to assay the anti-proliferation effect of Res in LoVo cells.The effect of Res on apoptosis in LoVo cells was also detected with flow cytometry.Then,RT-PCR and Western blot assay were employed to unveil the effect of Res on the ex-pression of BMP9 .The effect of BMP9 on the anti-pro-liferation of Res in LoVo cells was analyzed with crystal violet staining and flow cytometry too.Finally,the effect of Res on the expression of ALK2 and ALK3 was assayed with RT-PCR,and the inhibitor of ALK2 and ALK3 was used to figure out the possible mechanism of BMP9 on Res-induced proliferation inhibition in LoVo cells.Results Res apparently inhibited the prolifera-tion,arrested the cell cycle at S phase in LoVo and in-creased the percentage of apopotic cells in LoVo cells. Res increased the expression of mRNA and protein of BMP9 concentration dependently. Exogenous ex-pressed-BMP9 enhanced the anti-proliferation and ap-optosis inducing effects of Res in LoVo cells, but BMP9 knockdown decreased these effects of Res.Al-though Res had no apparent effect on increasing the phosphorylation of Smad1/5/8,it increased the ex-pression of ALK2 and ALK3 .Inhibition of ALK2 and ALK3 decreased the anti-proliferation effect of Res partly in LoVo cells.Conclusion Res is potent to in-hibit the proliferation of LoVo cells,Which may be mediated by up-regulating the expression of BMP9 and its receptor at least.

Effects of meloxicam on proliferation,migration and expression of PTEN of human colorectal cancer cells / 中国药理学通报

Aim To investigate the effects of meloxi-cam on the proliferation,migration and expression of PTEN of human colorectal cancer LoVo cells.Meth-ods The colony formation test was used to detect the effect of meloxicam on the proliferation of LoVo cells. The cell migration assay was applied to analyze the effect of meloxicam on LoVo cells activity.The RT-PCR assay was used to detect the effect of meloxicam on the mRNA expression of PCNA and PTEN gene. The western blot assay was applied to analyze the effects of meloxicam on the expression of PTEN pro-tein.The recombinant adenovirus and Annexin-V assay were used to testify the relationship between PTEN gene and anti-cancer effect of meloxicam.Results Compared with the control group,meloxicam could in-hibit the colony formation and PCNA protein expression of LoVo cells.At 48 h and 80 μmol·L -1 ,the expres-sion of PCNA protein was reduced to 61 .57% ± 2.81 %(T =7.086,P =0.01 9),the mRNA expres-sion of PTEN gene increased to 1 60.43% ±4.71 %(T=24.244,P =0.002),and the expression of PTEN protein increased to 1 52.63% ±3.33%(T =27.359, P =0.001 ).Results Annexin-V test indicated that the anti-cancer effect of meloxicam was associated with the up-regulated expression of PTEN.Conclusions Meloxicam can inhibit the proliferation and migration of LoVo cells by up-regulating the expression of PTEN.

Faecalibacterium prausnitzii Genomic DNA Enhances the Killing Activity of Peripheral Blood Mononuclear Cells against Human Colon Cancer LoVo Cells by Upregulating Th1 Immune Response / 胃肠病学

Background:Faecalibacterium prausnitzii(Fp)is a commensal intestinal bacterium that exhibits anti-inflammatory and immunomodulatory capacity in vivo and in vitro. It has been reported that Fp in intestinal lumen was reduced in patients with colorectal cancer,which might be a factor associated with cancer development. Aims:To investigate the effect and immunological mechanism of Fp and its genomic DNA(fDNA)on the killing activity of peripheral blood mononuclear cells (PBMCs)against human colon cancer LoVo cells. Methods:PBMCs derived from healthy adults were co-cultured in vitro with Fp,fDNA,or the digested fDNA(d-fDNA),respectively. Killing activity of PBMCs against LoVo cells was measured by MTT assay;concentrations of interferon-gamma(INF-γ),a Th1-type cytokine and interleukin-4(IL-4),a Th2-type cytokine in culture supernatant of PBMCs were determined by ELISA;and expressions of T-bet and GATA3,the transcription factors specific for Th1 and Th2 cells,were measured by real-time PCR. Results:Compared with the PBMCs not treated,fDNA could significantly enhance the killing activity of PBMCs against LoVo cells(P < 0. 05);meanwhile,it promoted IFN-γ secretion,up-regulated T-bet mRNA expression and inhibited IL-4 secretion and GATA3 mRNA expression in PBMCs(P < 0. 05). Similar effects were not observed in PBMCs treated with Fp and d-fDNA. Conclusions:fDNA enhances the killing activity of PBMCs against human colon cancer cells by up-regulating Th1 immune response.

Effect of Gambogic acid on proliferation and invasion in colon cancer LoVo cell line / 国际中医中药杂志

Objective To investigate the effects of gambogic acid on proliferation,invasion and protein expression of MMP-2 in LoVo human colon cancer cell line.Methods The CCK-8 assay was used to determine the anti-proliferation ability.Transwell chamber assay was used to study the invasion of LoVo colon cancer cells.Western Blot was applied to examine the protein expression of MMP-2.Results CCK-8 assay showed that after cells treated with 1,2,4 μmol/L GA for 24 h,48 h the inhibition rates were(0.16±0.11)％,(0.24±0.08)％,(0.58±0.01)％,(0.67±0.03)％,(0.79±0.01)％ and (0.27±0.05)％,(0.69±0.09)％,(0.85±0.01)％,(0.87±0.01)％,(0.89±0.01)％,and there had statistically significant differences between control group (0 μmo/L) with experimental group (P＜0.05).Gambogic acid can effectively inhibit LoVo cells invasion at a low-dose concentration,which the numbers of cells that digested the matrigel in control group (0 μmo/L)was 120.50± 8.69 and 69.83 ±4.75 in experimental group (1 μnol/L),P＜ 0.05).Western Blot revealed that Gambogic acid can down-regulate the expression of MMP-2,which the levels of down-regulating the expression of MMP-2 after treated with 1,2,4 umol/L GA were 48.67％,74.72％and 82.70％ (P＜0.05) Conclusion Gambogic acid can effectively inhibit the growth and invasion of the LoVo cells,which may be related with down-regulating the expression of MMP-2.

Effect of human tumor necrosis factor alpha-sercreting on proliferation of LOVO colon cancer cells / 局解手术学杂志

objective To observe the effects of co-culture of hTNFα-sercreting and human colon cancer cells LOVO on the proliferation of cancer cells. Methods The stable transfected hTNF-α/293 , mRNA of Hek-293 cell and protein expression were detected by RT-PCR and ELISA. The positive group was added hTNF-αfactor,and MTT assay was applied under the optical density 490 nm. Through human tumor cell proliferation inhibition experiment,the inhibitory effects on colon cancer cells ( LOVO) proliferation were observed. Results hTNF-α/293 cells and hTNF-α-positive group showed a significant lower A,which suggested that hTNF-α/293 cells and hTNF-α-positive group had significant inhibition on the proliferation of colon cancer cells. Conclusion The inhibition of hTNF-αsecreted by hTNF-α/293 cells on co-lon cancer cell proliferation shows significant dose-effect dependency,and hTNF-αexpresses a considerable inhibition on the colon cancer cell proliferation as positive drug.

Observation of metastasis of colon adenocarcinoma Iovo cells in vitro under laser scanning confocal microscope / 国际生物医学工程杂志

Objective To explore the feasibility of applying laser scanning confocal microscopy to investigate the metastatic characteristic of tumor cells in vitro. Methods The confocal microscope combined with transwell assay was employed to observe the invasion and migration of the metastatic colon cancer ]ovo cells when stimulated by IL-8. Results The results show that the number of invasive lovo cells in the stimulated group was much higher than the unstimulated group (P<0.05). The migration way of lovo cells observed under the confocal microscopy was ameba like movement. These findings suggest that the metastatic ability of lovo cells stimulated by IL-8 was enhanced. Conclusion The metastatic characteristic of colon adenocarcinoma lovo cells is similar to the chemotactic movement of leukocytes.

Effects of recombinant human growth hormone and fluorouracil on human colon carcinoma LOVO cells in vitro / 肠外与肠内营养

Objective: To investigate the effects of recombinant human growth hormone (rhGH) and 5-fluorouracil(5-Fu) on human colon carcinoma LOVO cells in vitro. Methods: The LOVO cells during exponential growth stage were harvested and divided into control group,GH group, 5-Fu group and GH + 5-Fu group. According to the dose of GH, the GH group was separated into two sub-groups(50 ng/mL and 100 ng/mL) and the GH +5-Fu group was separated into two sub-groups. With different concentrations of rhGH and/or 5-Fu , the cell survive rates were analyzed by MTT assay after 24 h , 48 h and 72 h and cell cycle and proliferation index (PI) were analyzed by flow cytometry after 24 h. Results: Compared with the control group, the survive rates in 5-Fu and GH +5-Fu groups were decreased significantly (P 0. 05). Conclusion-. rhGH does not stimulate the LOVO cells proliferation in vitro, and its use is safe when combined with 5-fluorouracil.

Up-expression of peroxisome proliferator-activated receptor-? and of retinoid X receptor-? enhances the inhibitory effect on tumor cells growth / 基础医学与临床

LOVO.PGZ significantly up-regulates the expression of PPAR? in MGC803 and LOVO cells,the expression of PPAR? was higher in combination group than PGZ alone(P

Effects of TNP-470 on proliferation, the cell cycle and apoptosis in human colon cancer Lovo cells / 中国药理学通报

AIM To study the effect of TNP 470 on proliferation, cell cycle and apoptosis in the cultured human colon cancer Lovo cells. METHODS The growth inhibition of TNP 470 on Lovo cells was evaluated by an MTT.assay The effect of TNP 470 on cell cycle and apoptosis measured by flow cytometry, and ultrastructural feature of Lovo cells were observed with electromicroscope. RESULTS TNP 470 inhibited the growth of Lovo cells. flow cytometry analysis showed that G 0/G 1 phase rate was increased but S phase rate was decreased. Apoptosis rate of TNP 470 treated group was significant high than that of control and typical chang of apoptosis in Lovo cells was observed. CONCLUSION TNP 470 can inhibit proliferation of Lovo cells, and this inhibition is associated with cell cycle block and apoptosis.

Comparative proteomics studies on the serum of patients of colorectal cancer, LoVo cell suspension and HT29 cell culture fluid by use of SELDI TOF mass spectrometry / 解放军医学杂志

Objective To compare the specific proteomics between LoVo cells suspension, HT29 cells culture fluid and the serum of patients oof colorectal cancer. Methods serum of colorectal cancer, LoVo cells suspension and HT29 cells culture fluid were detected by IMAC3 chip and proteinchip reader (CipherGen Inc., VS). Results A protein at M/Z value 11731D was checked out in the LoVo cell suspension and the patients′ serum. No similar biomarkers were found in HT29 cell suspension and serum of colorectal cancer. Conclusion There existed similar biomarkers between LoVo cell suspension and serum of colorectal cancer