TM9SF1 is implicated in promoting the proliferation and invasion of bladder cancer cells

Zhuo et al looked into the part of transmembrane 9 superfamily member 1(TM9SF1)in bladder cancer(BC),and evaluated if it can be used as a therapeutic target.They created a permanent BC cell line and tested the effects of TM9SF1 overexpression and suppression on BC cell growth,movement,invasion,and cell cycle advancement.Their results show that TM9SF1 can boost the growth,movement,and invasion of BC cells and their access into the G2/M stage of the cell cycle.This research gives a novel direction and concept for targeted therapy of BC.

Human epidermal growth factor receptor 2 expression level and combined positive score can evaluate efficacy of advanced gastric cancer

BACKGROUND Although treatment options for gastric cancer(GC)continue to advance,the overall prognosis for patients with GC remains poor.At present,the predictors of treatment efficacy remain controversial except for high microsatellite instability.AIM To develop methods to identify groups of patients with GC who would benefit the most from receiving the combination of a programmed cell death protein 1(PD-1)inhibitor and chemotherapy.METHODS We acquired data from 63 patients with human epidermal growth factor receptor 2(HER2)-negative GC with a histological diagnosis of GC at the Cancer Hospital,Chinese Academy of Medical Sciences between November 2020 and October 2022.All of the patients screened received a PD-1 inhibitor combined with chemotherapy as the first-line treatment.RESULTS As of July 1,2023,the objective response rate was 61.9%,and the disease control rate was 96.8%.The median progression-free survival(mPFS)for all patients was 6.3 months.The median overall survival was not achieved.Survival analysis showed that patients with a combined positive score(CPS)≥1 exhibited an extended trend in progression-free survival(PFS)when compared to patients with a CPS of 0 after receiving a PD-1 inhibitor combined with oxaliplatin and tegafur as the first-line treatment.PFS exhibited a trend for prolongation as the expression level of HER2 increased.Based on PFS,we divided patients into two groups:A treatment group with excellent efficacy and a treatment group with poor efficacy.The mPFS of the excellent efficacy group was 8 months,with a mPFS of 9.1 months after excluding a cohort of patients who received interrupted therapy due to surgery.The mPFS was 4.5 months in patients in the group with poor efficacy who did not receive surgery.Using good/poor efficacy as the endpoint of our study,univariate analysis revealed that both CPS score(P=0.004)and HER2 expression level(P=0.015)were both factors that exerted significant influence on the efficacy of treatment the combination of a PD-1 inhibitor and chemotherapy in patients with advanced GC(AGC).Finally,multivariate analysis confirmed that CPS score was a significant influencing factor.CONCLUSION CPS score and HER2 expression both impacted the efficacy of immunotherapy combined with chemotherapy in AGC patients who were non-positive for HER2.

Saposin C stimulates growth and invasion,activates p42/44 and SAPK/JNK signaling pathways of MAPK and upregulates uPA/uPAR expression in prostate cancer and stromal cells

Aim:To determine the effect of saposin C (a known trophic domain of prosaposin) on proliferation,migration and invasion,as well as its effect on the expression of urokinase plasmonogen activator (uPA),its receptor (uPAR) and matrix metalloproteinases (MMP)-2 and -9 in normal and malignant prostate cells.In addition,we tested whether saposin C can activate p42/44 and stress-activated protein kinase/c-Jun NH_2-terminal kinase (SAPK/JNK) signal transduction pathways of the mitogen-activated protein kinase (MAPK) superfamily.Methods:We employed West- ern blot analysis,phospho-specific antibodies,cell proliferation assay,reverse transcriptase-polymerase chain reaction, in vitro kinase assays and migration and invasion to determine the effect of saposin C on various biological behaviors of prostate stromal and cancer cells.Results:Saposin C,in a cell type-specific manner,upregulates uPA/uPAR and immediate early gene c-Jun expression,stimulates cell proliferation,migration and invasion and activates p42/44 and SAPK/JNK MAPK pathways in prostate stromal and cancer cells.Normal prostate epithelial cells were not responsive to saposin C treatment in the above studies.Conclusion:Saposin C functions as a multipotential modulator of diverse biological activities in prostate cancer and stromal cells.These results strongly suggest that saposin C functions as a potent growth factor for prostatic cells and may contribute to prostate carcinogenesis and/or the development of hormone-refractory prostate cancer.

Over-expression of LRIG3 Suppresses Growth and Invasion of Bladder Cancer Cells

The purpose of this study was to investigate the impact of leucine-rich repeats and immu- noglobulin-like domains 3 （LRIG3） on the biological features of bladder cancer cell lines. The plasmids of over-expressed LRIG3 and the blank plasmid serving as control were transfected into the bladder cancer cell lines, T24, EJ and BIU-87, and the expression levels of LRIG3 mRNA and protein were de- tected by using real-time PCR and Western blotting. The changes in the cell cycle and apoptosis were examined by using flow cytometry. The invasive ability was measured by Transwell assay, and CCK-8 assays were used to measure the proliferation of cells. As compared with the control group, the LRIG3 mRNA and protein expression levels in LRIG3 cDNA-transfected group were raised significantly （P〈0.05）. The average number of cells with up-regulated LRIG3 passing through the inserted filter was decreased significantly as compared with the control group （P〈0.05）. Up-regulation of LRIG3 also could inhibit proliferation and induce apoptosis of T24, EJ and BIU-87 cells. Except BIU-87, the T24 and EJ cells transfected with LIRG3 eDNA were arrested in G0/G1 phase compared to the control group （P〈0.05）. In conclusion, the over-expression of LRIG3 could influence the cell cycle and invasion, in- hibit proliferation and induce apoptosis in the three bladder cancer cell lines.

Suppression of cell growth and invasion by miR-205 in breast cancer

MicroRNAs (miRNAs ) 内长的、小、非编码的 RNA，它能够在 post-transcriptional 的 silencing 基因表示铺平。在这研究，我们报导 miR-205 是显著地在与匹配的正常的胸织物相比的胸肿瘤的 underexpressed。同样，包括 MCF-7 和 MDA-MB-231，乳癌房间行比非恶意的 MCF-10A 房间表示低级 miR-205。兴趣， miR-205 的宫外的表示显著地禁止房间增长和抛锚独立人士生长，以及房间侵略。而且， miR-205 被显示在一个动物模型压制肺转移。最后，西方的污点与试金表明的酶记者结合了那 ErbB3 和脉管的 endothelial 生长因素 A ( VEGF --一)是为 miR-205 的直接目标，并且这 miR-205-mediated 抑制通过和在 ErbB3 和 VEGF 的 3 鈥? untranslated 区域( 3 鈥? UTR )的通常认为的 miR-205 绑定地点的直接相互作用是可能的--一。一起，这些结果建议 miR-205 是在乳癌的肿瘤 suppressor。

Inhibitory Effect of MiR-449b on Cancer Cell Growth and Invasion through LGR4 in Non-Small-Cell Lung Carcinoma

Non-small-cell lung carcinoma （NSCLC） is one of the most frequently diagnosed malignancies worldwide. Previous studies have shown that microRNA-449b （miR-449b） functions as a tumor suppressor in many cancers. However, the role of miR- 449b in NSCLC is still unknown. In the present study, miR-449b was significantly down- regulated in NSCLC samples and cell lines. Bioinformatics analysis revealed that 3＇-UTR region of leucine rich repeat containing G protein-coupled receptor 4 （LGR4） mRNA had putative complementary sequences to miR-449b, which was further confirmed by the luciferase assay. Western blotting showed that restoration of miR-449b in NSCLC cells decreased the expression of LGR4. Interestingly, over-expression of miR-449b inhibited growth and invasion of NSCLC cells in vitro. Furthermore, ectopic expression of LGR4 reversed miR-449b-suppressed proliferation and invasion of NSCLC cells. Therefore, the data of the present study demonstrate that miR-449b inhibits tumor cell growth and invasion by targeting LGR4 in NSCLC.

shRNA-interfering LSD1 inhibits proliferation and invasion of gastric cancer cells via VEGF-C/PI3K/AKT signaling pathway

BACKGROUND Histone Lysine Specific Demethylase 1(LSD1)is the first histone demethylase to be discovered,which regulates various biological functions by making lysine of histone H3K4,H3K9 and non-histone substrates demethylated.Abnormal regulation of LSD1 is closely related to the occurrence and development of gastric cancer.The change of LSD1 expression level plays an important role in the proliferation and metastasis of gastric cancer cells.The study of its function and mechanism may provide a theoretical basis for early diagnosis and targeted therapy of gastric cancer.AIM To investigate the effect of downregulation of lysine-specific demethylase 1(LSD1)expression on proliferation and invasion of gastric cancer cells and the possible regulatory mechanisms of the VEGF-C/PI3K/AKT signaling pathway.METHODS The LSD1-specific short hairpin RNA(shRNA)interference plasmid was transiently transfected,and expression of LSD1 was downregulated.The cell proliferation ability of LSD1 was observed by CCK-8 assay after downregulating expression of LSD1.Transwell invasion assay was used to observe the change of cell invasion ability after downregulating expression of LSD1.Expression of phosphorylated phosphoinositide 3-kinase(p-PI3K),PI3K,p-AKT,AKT,vascular endothelial growth factor receptor(VEGFR)-3,matrix metalloproteinase(MMP)-2 and MMP-9 in each group was detected by Western blotting.RESULTS The cell proliferation ability of transiently transfected LSD1-shRNA interference plasmid group was significantly lower than that of the control group(P<0.05).Transwell invasion assay showed that the number of cells across the membrane of the LSD1-shRNA transfection group(238.451±5.216)was significantly lower than that of the control group(49.268±6.984)(P<0.01).Western blotting showed that expression level of VEGF-C,p-PI3K,PI3K,p-AKT,AKT,VEGFR-3,MMP-2 and MMP-9 in the LSD1-shRNA group was significantly lower than that in the control group(P<0.05).CONCLUSION Downregulation of LSD1 expression inhibits metastatic potential of gastric cancer cells,and VEGF-C-mediated activation of PI3K/AKT signaling pathway,which may be an important mechanism for inhibiting lymph node metastasis in gastric cancer cells.

Expression of Nerve Growth Factor and Hypoxia Inducible Factor-1α and Its Correlation with Angiogenesis in Non-Small Cell Lung Cancer

Summary： In order to investigate the expression of nerve growth factor （NGF） and hypoxia inducible factor-1α （HIF-1α） and its correlation with angiogenesis in non-small cell lung cancer （NSCLC）, paraffin-embedded tissue blocks from 20 patients with NSCLC were examined. Twenty corresponding para-cancerous lung tissue specimens were obtained to serve as a control. The expression of NGF, HIF-1α, and vascular endothelial growth factor （VEGF） in the NSCLC tissues was detected by using immunohistochemistry. The microvascular density （MVD） was determined by CD31 staining. The resuits showed that the expression levels ofNGF, HIF-1α and VEGF in the NSCLC tissues were remarkably higher than those in the para-cancerous lung tissues （P〈0.05）. There was significant difference in the MVD between the NSCLC tissues （9.19±1.43） and para-cancerous lung tissues （2.23±1.19） （P〈0.05）. There were positive correlations between NGF and VEGF, between HIF-1α and VEGF, and between NGF and HIF-1α in NSCLC tissues, with the spearman correlation coefficient being 0.588, 0.519 and 0.588, respectively. In NSCLC tissues, the MVD had a positive correlation with the three factors （P〈0.05）. Theses results suggest that NGF and HIF-1α are synergically involved in the angiogenesis of NSCLC.

Nexus of signaling and endocytosis in oncogenesis driven by non-small cell lung cancer-associated epidermal growth factor receptor mutants

Epidermal growth factor receptor(EGFR) controls a wide range of cellular processes, and aberrant EGFR signaling as a result of receptor overexpression and/or mutation occurs in many types of cancer. Tumor cells in non-small cell lung cancer(NSCLC) patients that harbor EGFR kinase domain mutations exhibit oncogene addiction to mutant EGFR, which confers high sensitivity to tyrosine kinase inhibitors(TKIs). As patients invariably develop resistance to TKIs, it is important to delineate the cell biological basis of mutant EGFR-induced cellular transformation since components of these pathways can serve as alternate therapeutic targets to preempt or overcome resistance. NSCLC-associated EGFR mutants are constitutively-active and induce ligandindependent transformation in nonmalignant cell lines. Emerging data suggest that a number of factors are critical for the mutant EGFR-dependent tumorigenicity, and bypassing the effects of TKIs on these pathways promotes drug resistance. For example, activation of downstream pathways such as Akt, Erk, STAT3 and Src is critical for mutant EGFR-mediated biological processes. It is now well-established that the potency and spatiotemporal features of cellular signaling by receptor tyrosine kinases such as EGFR, as well as the specific pathways activated, is determined by the nature of endocytic traffic pathways through which the active receptors traverse. Recent evidence indicates that NSCLCassociated mutant EGFRs exhibit altered endocytic trafficking and they exhibit reduced Cbl ubiquitin ligasemediated lysosomal downregulation. More recent work has shown that mutant EGFRs undergo ligand-independent traffic into the endocytic recycling compartment, a behavior that plays a key role in Src pathway activation and oncogenesis. These studies are beginning to delineate the close nexus between signaling and endocytic traffic of EGFR mutants as a key driver of oncogenicprocesses. Therefore, in this review, we will discuss the links between mutant EGFR signaling and endocytic properties, and introduce potential mechanisms by which altered endocytic properties of mutant EGFRs may alter signaling and vice versa as well as their implications for NSCLC therapy.

Effect of h GC-MSCs from human gastric cancer tissue on cell proliferation, invasion and epithelial-mesenchymal transition in tumor tissue of gastric cancer tumor-bearing mice

Objective: To study the effect of h GC-MSCs from human gastric cancer tissue on cell proliferation, invasion and epithelial-mesenchymal transition in tumor tissue of gastric cancer tumor-bearing mice. Methods: BABL/c nude mice were selected as experimental animals and gastric cancer tumor-bearing mice model were established by subcutaneous injection of gastric cancer cells, randomly divided into different intervention groups. h GC-MSCs group was given different amounts of gastric cancer cells for subcutaneous injection, PBS group was given equal volume of PBS for subcutaneous injection. Then tumor tissue volume were determined, tumor-bearing mice were killed and tumor tissues were collected, m RNA expression of proliferation, invasion, EMT related molecules were determined. Results: 4, 8, 12, 16, 20 d after intervention, tumor tissue volume of h GC-MSCs group were significantly higher than those of PBS group and the more the number of h GC-MSCs, the higher the tumor tissue volume; m RNA contents of Ki-67, PCNA, Bcl-2, MMP-2, MMP-7, MMP-9, MMP-14, N-cadherin, vimentin, Snail and Twist in tumor tissue of h GC-MSCs group were higher than those of PBS group, and m RNA contents of Bax, TIMP1, TIMP2 and E-cadherin were lower than those of PBS group. Conclusions: h GC-MSCs from human gastric cancer tissue can promote the tumor growth in gastric cancer tumor-bearing mice, and the molecular mechanism includes promoting cell proliferation, invasion and epithelial-mesenchymal transition.

Activation of Rac1-PI3K/Akt is required for epidermal growth factorinduced PAK1 activation and cell migration in MDA-MB-231 breast cancer cells

Epidermal growth factor （EGF） may increase cell motility, an event implicated in cancer cell invasion and metastasis. However, the underlying mechanisms for EGF-induced cell motility remain elusive. In this study, we found that EGF treatment could activate Ras-related C3 botulinum toxin substrate 1 （Racl）, PI3K/Akt and p21- actived kinase （PAK1） along with cell migration. Ectopic expression of PAK1 K299R, a dominant negative PAK1 mutant, could largely abolish EGF-induced cell migration. Blocking PI3K/Akt signalling with LY294002 or Akt siRNA remarkably inhibited both EGF-induced PAK1 activation and cell migration. Furthermore, expression of dominant-negative Racl （T17N） could largely block EGF-induced PI3K/Akt-PAK1 activation and cell migration. Interestingly, EGF could induce a significant production of ROS, and N-acetyl-L-cysteine, a scavenger of ROS which abolished the EGF-induced ROS generation, cell migration, as well as activation of PI3K/Akt and PAK, but not Racl. Our study demonstrated that EGF-induced cell migration involves a cascade of signalling events, including activation of Racl, generation of ROS and subsequent activation of PI3K/Akt and PAK1.

Relationship between VEGF,EGF and Invasion,Metastasis of Gastric Cancer Cells

Objective： To investigate the relationship between expression of vascular endothelial growth factor （VEGF）, epidermal growth factor （EGF） and the biological properties of gastric cancer cells such as invasion and metastasis. Methods： RT-PCR was performed to semi-quantitatively detect the mRNA expressions of EGF, EGFR, VEGF and VEGFR in four kinds of gastric cancer cell lines BGC823, MGCS03, HGC27 and SGC7901, which were classified by their differentiation degree in our experiment. We obtained cell line growth curves from MTT assays. The migration of gastric cancer cells was observed under inverted phase contrast microscope. The changes of invasion and adhesion were detected by a Transwell assay. Results： The growth rates slowed down sequentially in MGC803, HGC27, BGC823 and SGC7901（P〈 0.05）. The ability of migration, invasion and adhesion were reduced sequentially, and the difference was significant. The expressions of EGF, EGFR, VEGF and VEGFR were significantly stronger in MGCS03 and HGC27 than in BGC823 and SGC7901 cells, and the difference was statistically significant（P〈0.05）. Conclusion： The expressions of VEGF and EGF had close relationship with the properties of migration, adhesion and invasion of gastric cancer cells in vitro. Thus, targeting VEGF and EGF may be a potential therapeutic strategy for inhibiting peritoneal metastasis of gastric cancer.

The Effects of Liquor Spirits on RNA Pol III Genes and Cell Growth of Human Cancer Lines

Alcohol consumption is a major health issue and associated with human cancers, such as liver and breast cancers. Alcohol was classed as carcinogen to human by IARC. We have performed in vivo and in vitro studies which demonstrate that diluted ethanol promotes cell proliferation and transformation and tumor formation. Consumption of liquor spirits (white wines) is a popular behavior. However, it is unclear whether liquor spirits affect cellular phenotypes of human cancers. At present study, we used diluted ethanol and liquor spirits (Sample #1 and Sample #2) to determine the changes in RNA polymerase III-dependent gene (Pol III gene) transcription, cell growth and colony formation in the different human cancer lines. The results indicate that low concentration of ethanol increases RNA Pol III gene transcription and rate of cell growth. However, both liquor spirits (Sample #1 and Sample #2) inhibit the activity of RNA Pol III genes and repress cell proliferation of the cancer lines, compared to diluted ethanol. The liquor spirits reduce the rate of colony formation of human breast cancer cells and esophageal carcinoma cells. The inhibitions of the liquor spirits to RNA Pol III genes, cell growth and colony formation are in a dose-dependent manner. These new findings suggest that the liquor spirits contain some active components to repress Pol III gene transcription and cell growth caused by ethanol in different human cancer cells.

PBX3 promotes migration and invasion of colorectal cancer cells via activation of MAPK/ERK signaling pathway

AIM:To investigate the role of pre-B-cell leukemia homeobox(PBX)3 in migration and invasion of colorectal cancer(CRC)cells.METHODS:We detected PBX3 expression in five cell lines and surgical specimens from 111 patients with CRC using real-time reverse transcription-polymerase chain reaction.We forced expression of PBX3 in low metastatic HT-29 and SW480 cells and knocked down expression of PBX3 in highly metastatic LOVO and HCT-8 cells.Wound healing and Boyden chamber assays were used to detect cell migration and invasionafter altered expression of PBX3.Western blot was performed to detect the change of signaling molecule ERK1/2 following PBX3 overexpression.RESULTS:High level of PBX3 expression was correlated with the invasive potential of CRC cells,and significantly associated with lymph node invasion(P=0.02),distant metastasis(P=0.04),advanced TNM stage(P=0.03)and poor overall survival of patients(P<0.05).Ectopic expression of PBX3 in low metastatic cells was shown to promote migration and invasion,while inhibited PBX3 expression in highly metastatic cells suppressed migration and invasion.Furthermore,upregulation of phosphorylated extracellular signal-regulated kinase(ERK)1/2 was found to be one of the targeted molecules responsible for PBX3-induced CRC cell migration and invasion.CONCLUSION:PBX3 induces invasion and metastasis of CRC cells partially through activation of the MAPK/ERK signaling pathway.

Effects of hepatocyte growth factor/scatter factor on the invasion of colorectal cancer cells in vitro

AIM: Hepatocyte growth factor (HGF) is a multifunctional growth factor which has pleiotrophic biological effects on epithelial cells, such as proliferation, motogenesis, invasiveness and morphogenesis. There are few reports about the role of HGF played in the colorectal cancer invasion.In the present study, we tried to investigate the possible mechanism of HGF involved in the invasion of colorectal cancer cells in vitro.METHODS: Matrigel migration assay was used to analyze the migrational ability of Caco-2 and Colo320 in vitro.We detected the mRNA expressive levels of MMP-2,MMP-9 and their natural inhibitors TIMP-1, TIMP-2 in Caco-2 cells by reverse-transcription polymerase chain reaction (PCR) technique.RESULTS: After 48 h incubation, there were notable differences when we compared the migrational numbers of Caco-2 cells in the group of HGF and PD98059 (the inhibitor of p42/p44MAPK) with the control (104.40±4.77vs 126.80±5.40, t= 7.17, P= 0.002＜0.01; 104.40±4.77vs 82.80±4.15, t= 7.96, P= 0.001＜0.01). The deviation between the HGF and PD98059 was significant (P＜0.01).Compared with controls, MMP-2 and MMP-9 mRNA expres-sions were up-regulated by HGF (0.997±0.011 vs 1.207±0.003, t= 35.002,P= 0.001＜0.01; 0.387±0.128 vs 0.971±0.147, t= 106.036, P= 0.0000＜0.01, respectively);compared with controls, TIMP-1, TIMP-2 mRNA expressions were increased by PD98059 (1.344±0.007 vs 1.905±0.049,t= 17.541, P= 0.003＜0.01; 1.286±0.020 vs 1.887±0.022,t = 24.623, P = 0.002＜0.01, respectively).CONCLUSION: HGF promoted Caco-2 migration mainly by p42/p44MAPK pathway; HGF/SF stimulated the expression of MMP-2, MMP-9 in Caco-2 and enabled tumoral cells to damage the ECM and reach the distant organ and develop metastasis; HGF played the function of promoted-invasion and promoted-metastasis, in which cellular selection was possible.

Direct interaction between Rab5a and Rab4a enhanced epidermal growth factor-stimulated proliferation of gastric cancer cells

BACKGROUND Gastric cancer(GC)is one of the leading causes of cancer-related death worldwide.Although targeted therapies such as antibodies against human epidermal growth factor receptor 2 or vascular endothelial growth factor receptor 2 have been widely used in the treatment of metastatic cancer,the overall outcomes are poor.Therefore,elucidation of the mechanism underlying cancer progression is important to improve prognosis.Overexpression of the Rab5a gene has been confirmed to correlate with tumorigenesis of many cancers,but the mechanism underling,especially of GC,is still unclear.AIM To investigate the effects of Rab5a overexpression on the tumorigenesis of GC.METHODS First,the expression levels of Rab5a and Rab4a in primary tumorous tissues of GC patients diagnosed between 2015 and 2018 were analyzed.Then we constructed HGC-27 cell lines overexpressing green fluorescent protein-Rab5a or red fluorescent protein-Rab4a and investigated the interaction between Rab5a or Rab4a using Western blotting,co-immunoprecipitation,confocal microscopy,and colocalization analysis.Finally,epidermal growth factor-stimulated proliferation of these cell lines was analyzed using cell counting kit-8 cell viability assay.RESULTS Compared with normal gastric tissues,the expression levels of Rab5a and Rab4a increased progressively both in paracancerous tissues and in advanced cancerous tissues.Epidermal growth factor could promote the proliferation of HGC-27 cells,especially Rab5a-overexpressing HGC-27 cells.Notably,Rab5a and Rab4a cooverexpression promoted the proliferation of HGC-27 cells to the greatest extent.Further analysis identified a direct interaction between Rab5a and Rab4a in HGC-27 cells.CONCLUSION Co-overexpression of Rab5a and Rab4a in GC may promote the endosomal recycling of epidermal growth factor receptor,which in turn contributes to poor prognosis and tumor progression in GC patients.Inhibition of Rab5a or Rab4a expression might be a promising therapy for refractory GC.

Integrin-linked kinase in gastric cancer cell attachment,invasion and tumor growth

AIM:To investigate the effects of integrin-linked kinase(ILK)on gastric cancer cells both in vitro and in vivo.METHODS:ILK small interfering RNA(siRNA)was transfected into human gastric cancer BGC-823 cells and ILK expression was monitored by real-time quantitative polymerase chain reaction,Western blotting analysis and immunocytochemistry.Cell attachment,proliferation,invasion,microfilament dynamics and the secretion of vascular endothelial growth factor(VEGF) were also measured.Gastric cancer cells treated with ILK siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed.RESULTS:Both ILK mRNA and protein levels were significantly down-regulated by ILK siRNA in human gastric cancer cells.This significantly inhibited cell attachment,proliferation and invasion.The knockdown ofILK also disturbed F-actin assembly and reduced VEGF secretion in conditioned medium by 40%(P<0.05).Four weeks after injection of ILK siRNA-transfected gastric cancer cells into nude mice,tumor volume and weight were significantly reduced compared with that of tumors induced by cells treated with non-silencing siRNA or by untreated cells(P<0.05).CONCLUSION:Targeting ILK with siRNA suppresses the growth of gastric cancer cells both in vitro and in vivo.ILK plays an important role in gastric cancer progression.

Effect of ligand troglitazone on peroxisome proliferator-activated receptor γ expression and cellular growth in human colon cancer cells

AIM: To investigate the effect of troglitazone on pe- roxisome proliferator-activated receptor γ (PPARγ) expression and cellular growth in human colon cancer HCT-116 and HCT-15 cells and to explore the related molecular mechanism. METHODS: Human colon cancer HCT-116 and HCT-15 cells cultured in vitro were treated with troglitazone. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were employed to detect the effect of troglitazone on PPARγ expression. The proliferative activity was determined by MTT assay, cell cycle and apoptosis were detected by ? ow cytometry. Apoptosis- related genes, cell cycle regulatory genes and p53 were examined by RT-PCR and Western blot respectively. RESULTS: The expression of PPARγ in colon cancer HCT-116 and HCT-15 cells was up-regulated by troglitazone. Troglitazone inhibited proliferation, induced apoptosis and cell cycle G1 arrest in colon cancer cells. Troglitazone induced p53 expression in HCT-116 cells, but not in HCT-15 cells. The down-regulation of survivin and bcl-2 was found in both cell lines and up-regulation of bax was found only in HCT-116 cells, being consistent with growth inhibition in HCT-116 cells but not in HCT-15 cells. Troglitazone increased expression of p21WAF1/CIP1 (p21), p27KIP1 (p27) and reduced cyclin D1 in HCT-116 cells while only a minor decrease of cyclin D1 was found in HCT-15 cells. CONCLUSION: Troglitazone is an inductor of PPARγin colon cancer cells and inhibits PPARγ-dependently proliferation, which may attribute to cell cycle G1 arrest and apoptosis in colon cancer cells. Troglitazone may induce p53-independent apoptosis and p53- dependent expression of p21 and p27. Depending on cell background, different activation pathways may exist in colon cancer cells.

Mechanism of SNHG12 in Regulating Human Angiostatin Binding Protein Through MicroRNA497 in the Migration and Invasion of Human Lung Cancer Cells

Objective:To investigate the effect of small nucleolar host gene 12(SNHG12)on the migration and invasion of human lung cancer cells by regulating human angiostatin binding protein through microribonucleic acid(microRNA)-497.Methods:A549,H1299,and PC9 cells were cultured in Roswell Park Memorial Institute(RPMI)-1640 medium containing 10%fetal bovine serum,and human bronchial epithelial(HBE)cells were cultured in Dulbecco’s modified eagle medium(DMEM)containing 10%fetal bovine serum.The incubator conditions were as follows:saturated humidity,37℃,and 5%carbon dioxide(CO2).Results:The gene expressions of small nucleolar host gene 12(SNHG12)in HBE,A549,H1299,and PC9 were 1.00±0.02,5.61±0.42,3.78±0.29,and 3.51±0.23,respectively.The gene expressions of microRNA-497HBE,A549,H1299,and PC9 were 1.00±0.13,0.21±0.04,0.35±0.05,and 0.37±0.06,respectively,with P<0.05.The microRNA-497 gene expression and cell apoptosis rate in the microRNA-497 group and the microRNA-497+pcDNA3.1 group were significantly higher than those in the miR-NC group,whereas the A value and cell invasion number were significantly lower than those in the miR-negative control(NC)group,with P<0.05.Compared with the microRNA-497+pcDNA3.1 group,the microRNA-497 gene expression and cell apoptosis rate in the microRNA-497+SNHG12 group were significantly lower,whereas the A value and cell invasion number were significantly higher,with P<0.05.Conclusion:SNHG12 can inhibit the migration and invasion of human lung cancer cells by regulating human angiostatin binding protein through microRNA-497.

Inhibitory Effects of AURKB Gene on Apoptosis and Cancer Cell Growth in HCT 116 Cells

[Objectives]To explore the inhibitory effect of AURKB gene in apoptosis and cancer cell growth in HCT 116 cells.[Methods]The in vitro cytology studies were carried out to confirm the expression of the AURKB gene in HCT 116 cells and make clear its role in cell activity,cell cycle control and apoptosis,and investigate the effect of AURKB gene in colorectal cancer(CRC).Quantitative reverse transcription/polymerase chain reaction(PCR)analysis and immunofluorescence(IF)staining for markers of AURKB gene were used to examine the effect of AURKB on HCT 116.The AURKB gene target was examined using western blot analysis.In addition,inhibition of AURKB expression was examined using RNA interference(RNAi)on HCT 116 cells in vitro.[Results]HCT 116 cells infected with AURKB shRNA virus suppressed expression of AURKB in vitro.AURKB gene knockdown HCT 116 cells showed reducing cell apoptosis in vitro.Finally,it demonstrated that AURKB function can induce apoptosis of HCT cells.[Conclusions]AURKB is a key regulator of colorectal cancer.AURKs are potential novel molecular targets for the prevention of cancer cell proliferation.