Results: A total of 20 studies comprised of 5876 individuals were eligible. There was no heterogeneity for CRC, but adenoma and advanced adenoma harbored considerable heterogeneity influenced by risk classification and various

detection markers. Stratification analysis by risk classification showed that multiple markers had a high diagnostic value for the high-risk subgroups of both CRC [sensitivity: 0.759(0.711–0.804), specificity: 0.883(0.846–0.913), AUC = 0.906] and advanced adenoma [sensitivity: 0.683(0.584–0.771), specificity: 0.918(0.866–0.954), AUC = 0.946] but not for the average-risk subgroups of either. In the methylation subgroup, sDNA had significantly higher diagnostic selleck chemicals llc value for CRC [sensitivity: 0.753(0.685–0.812), specificity: 0.913(0.860–0.950), AUC = 0.918] and advanced adenoma [sensitivity: 0.623(0.527–0.712),

specificity: 0.926(0.882–0.958), AUC = 0.910] compared to the mutation subgroup. There was no significant heterogeneity among studies for subgroup analysis. Conclusion: Multiple markers’ sDNA testing had strong diagnostic significance for CRC and advanced adenoma in high-risk subjects. Methylation makers have more diagnostic value than mutation markers. Key Word(s): 1. stool DNA test; 2. colorectal cancer; 3. adenoma; 4. diagnosis; Presenting Author: ZIJUN LI Additional Authors: WENJING NIE Corresponding Author: ZIJUN LI Affiliations: Guangdong General Hospital Objective: To investigate the distribution characteristics and clinical significance of lipid Trichostatin A molecular weight metabolism in patients with colorectal cancer. Methods: Total cholesterol (CHOL),

triglyceride (TGIG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A1 (ApoA1) and apolipoprotein B100 (ApoB100) were measured in patients newly diagnosed with colorectal Progesterone cancer in Guangdong General Hospital during 2001–2010, and all the data were analyzed. Results: A total of 1557 patients were included for final analysis, of whom 807 were in 2001–2005 group and 750 in 2006–2010 group. Both CHOL[(4.56 ± 1.07 vs. 4.40 ± 1.08)mmol/ L], TGIG[(1.12 ± 0.57 vs. 1.05 ± 0.70)mmol/ L], LDL-C[(2.78 ± 0.91 vs. 2.57 ± 0.92)mmol/ L], ApoB100[(0.72 ± 0.25 vs. 0.76 ± 0.20)mmol/ L] level in 2006–2010 group were higher than that of 2001–2005 group. And there were no statistical differences in the concentration of HDL-C, ApoA1 between the two groups. The rate of patients with evaluated levels of CHOL (26.5% vs. 21.7%), LDL-C (24.0% vs. 17.0%), and decreased levels of ApoA1 (72.3% vs. 67.5%) in 2006–2010 group were higher than that of 2001–2005 group (p < 0.05). No statistical differences were found between these two groups in the level of TRIG, HDLC, ApoB100. Conclusion: Patients with colorectal cancer often develop lipid metabolism abnormalities. Patients in 2006–2010 had higher level of blood lipid than those in 2001–2005.

16, 17 Protein extracts from human liver tumors and from healthy patients were obtained from OriGene (Rockville, MD). Liver tumors were induced in wild-type (WT) and Little mice via diethylnitrosoamine (DEN) tumor liver

induction as described.5 For FXR agonist treatment experiment, 8-week-old mice were injected with FXR agonist GW4064 intraperitoneally (30 mg/kg body weight). Autophagy Compound Library chemical structure Control mice were injected with vehicle (corn oil). Nuclear and cytoplasmic extract isolation and western blot analysis were performed as described our previous publications.18, 19 A typical picture of the quality of the separation of cytoplasmic and nuclear proteins is shown in Supporting Fig. 2. Total RNA from liver tissues or Hep3B2 cells was extracted with an RNeasy Mini Kit (Qiagen,

Germantown, MD) according to the manufacturer’s instructions. Complementary DNA was synthesized using SuperScript III First-strand (Invitrogen) and random primer hexamers. The primer sequences used in the studies are presented in the Supporting Information. Chromatin immunoprecipitation assay (ChIP) was performed as described5, 18 using the ChIP-IT kit (Active Motif, Carlsbad, CA). Electrophoretic mobility shift assay was performed as described.19 Antibodies to FXR (C20 and H130), gankyrin, C/EBPβ (C19), C/EBPα (A144), cdk4, cdc2, cyclin D3, Rb, p53, and HDAC1 (H-51) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies to acetyl-histone H3 (Lys9) and histone H3 trimethyl Lys9 were obtained from Abcam (Cambridge, MA). Monoclonal anti–β-actin antibody was from Sigma (St. Louis, MO). The bromodeoxyuridine Selleckchem Y-27632 (BrdU) uptake

assay kit was obtained from Invitrogen (Carlsbad, CA). Co-immunoprecipitation studies were performed using TrueBlot reagents as described.5, 19 Hepa 1-6 cells were transduced with the shRNA-expressing lentivirus (Sigma-Aldrich, St. Louis, MO), and stable cell lines were generated by selection with puromycin for 2 weeks. For in vivo silencing experiments, 3-month-old mice were injected via the tail vein with C/EBPβ siRNA or nontarget siRNA (50 selleck chemicals μg of siRNA from Dharmcon complexed with in vivo-jet PEI, N/P ratio of 6 per mouse). FXR/SHP KO mice have hepatobiliary dysfunctions, including increased liver proliferation16 and development of liver cancer at age of 12 months (Anakk et al., submitted for publication). Because gankyrin-mediated elimination of C/EBPα is one of the key events in the development of liver cancer,5 we examined whether this pathway is activated in the livers of FXR/SHP KO mice. Figure 1A shows a typical liver of a 17-month-old FXR/SHP KO mouse with advanced cancer. BrdU uptake confirmed that liver proliferation was increased in these animals (Fig. 1B). Because C/EBPα needs to be phosphorylated at S193 by cdc2 and cdk4 to be degraded by gankyrin,5 we examined the expression of C/EBPα, gankyrin, cdc2, and cdk4 in livers of FXR/SHP KO mice. Figure 1C shows that gankyrin was elevated in the livers of FXR/SHP KO mice.

We performed stratification analyses on cancer type (divided into digestive system cancers and other cancers). For digestive system cancers, we further separated Asians and Caucasians. Meta regression was used to illustrate potential reasons of

between-study heterogeneity. Egger’s test and Mitomycin C order inverted funnel plots were utilized to provide a diagnosis of publication bias (linear regression asymmetry test65). All analyses were performed using Stata version 9.2 software (Stata, College Station, TX, USA). All statistical evaluations were made assuming a two-sided test with a significance level of 0.05, unless stated otherwise. The characteristics of the selected studies are listed in Table 1. The distribution of genotypes in the controls was consistent with the Hardy–Weinberg equilibrium for all selected studies, except for three studies for −765G>C,33,37,48 two studies for −1195G>A,19,29 and two studies for 8473T>C.46,59 When we assumed that the OR for an allelic genetic association was 1.2, only one

study achieved a statistical power greater than 80%.61 Only learn more two studies had a detailed dominant genotype frequency, so we extracted the data only for the dominant model.18,41 The evaluation of the association between these three polymorphisms and cancer risk is presented in Table 2. Overall, the variant A allele of COX-2−1195G>A can significantly increase the risk of cancer in all of SPTLC1 the tested models (GA vs GG: OR, 1.18; 95% CI, 1.07–1.29; P = 0.267 for the heterogeneity test; AA vs GG: OR, 1.35; 95% CI, 1.14–1.60; P = 0.010 for the heterogeneity test; dominant model, GA/AA vs GG: OR, 1.29; 95% CI, 1.18–1.41; P = 0.113 for the heterogeneity test; recessive model, AA vs GG/GA: OR, 1.22; 95% CI, 1.10–1.34; P = 0.002 for the heterogeneity test). However, for COX-2−765G>C and 8473T>C, no significant associations between the polymorphisms and risk of cancer were

observed. We then evaluated the effect of the three polymorphisms by specific tumor types. As shown in Table 2and Figure 1, we found that −1195G>A can significantly increase the risk of digestive system cancers in all tested models (GA vs GG: OR, 1.23; 95% CI, 1.11–1.37; P = 0.278 for the heterogeneity test; AA vs GG: OR, 1.55; 95% CI, 1.28–1.88; P = 0.045 for the heterogeneity test; dominant model, GA/AA vs GG: OR, 1.36; 95% CI, 1.23–1.51; P = 0.149 for the heterogeneity test; recessive model, AA vs GG/GA: OR, 1.32; 95% CI, 1.16–1.51; P = 0.016 for the heterogeneity test), whereas the increased risk was not evaluated for the ‘other cancers’ group.

Additional Supporting Information may be found in the online version of this article. “
“The retinoid X receptor a (RXRα; NR2B1), the most highly expressed RXR isoform in liver, plays a central role in regulating bile acid, cholesterol, fatty acid, steroid Autophagy inhibitor libraries and xenobiotic metabolism and homeostasis. Studies indicated that post-translational modification of RXRα, in particular, inflammation-induced phosphorylation of several sites, can lead to further post-translational modification (e. g., ubiquitination

and SUMOylation), as well as altered RXRa stability and subce l l ular localization. We have found that inflammation-induced reduction in RXRα nuclear quantities involves JNK-dependent phosphorylation at Ser260.Details regarding the fate, induction, localization and function of RXRα where Ser260 is phosphorylated are poorly understood, due to the lack of specific detecting reagents, cell

lines and mouse models. To begin to address these important issues, we developed and characterized an Metformin anti-pRXRα Ser260 antibody (p260 Ab) and verified its specificity and sensitivity with shRNA knockdown, immunoblotting and confocal immunofluorescence assays in both human and mouse models. The phosphorylation of RXRα Ser260 is significantly increased in both nuclear and cytoplasmic compartments of IL-1 β-treated Huh-7 cells and LPS-treated mouse liver, with a novel

finding of a submembrane localization at baseline which is increased in response to inflammation. Moreover, the JNK Florfenicol inhibitor, SP600125, inhibits IL-1 –β-induced upregulation of pRXRα Ser260 in Huh-7 cells, suggesting that JNK is a necessary upstream kinase involved in RXRα Ser260 phosphorylation. Initial explorations with confocal immunofluorescence microscopy and co-immunoprecipitation (Co-IP) assays identified submembrane phospho-RXRα interactions with the submembrane protein β-catenin. Moreover, Co-IP and immunofluorescence assays revealed that inflammation increases the interaction between phospho-RXRα and β-catenin in IL-1 β treated Huh-7 cells and LPS treated mouse livers in both cytoplasmic and subplasma membrane locales extend our knowledge of the potential biological roles played by RXRα species. We conclude that inflammatory stimuli induce JNK-dependent RXRα Serine260 phosphorylation, the interaction between p-catenin and RXRα, and the subcellular redistribution of RXRα, including heretofore novel cytoplasmic and submembrane locales. Disclosures: The following people have nothing to disclose: Hong Tang, Zhining Den, Astrid Kosters, Daniel A. Moore, Saul J.

1%). Single and multiple infections were found in 42.38% and 16.54% of the samples, respectively. The most common multiple infection was of ToMV, PVY or both. These results show that the percentage of infected plants and plots in open field cultivation is very high in this region and the origin of the seed is an important factor in the incidence of virus Volasertib cell line infection. In this respect, preventive measures, including virus-free

seed, resistant cultivars and improved cultural practices, could reduce the incidence of virus infection. “
“The distinguished plant cell wall component referred to as hydroxyproline-rich glycoproteins (HRGPs) exists in two forms: soluble in the symplast and insoluble in the apoplast. Insolubilization of HRGPs in cell walls through oxidative cross-linking which is elicited by stress represents a characteristic feature exhibited by two classes of HRGPs, namely, extensins and proline/HRGPs. Cross-linking of these HRGPs is an important process to strengthen the ABT-737 datasheet cell walls that contributes to plant defence reactions. In this review, the available information on these proteins is analysed with respect

to their roles in host-pathosystems and the various techniques applied for their characterization. Future prospects on strengthening of cell walls through gene regulation and transgenic approaches are also addressed. “
“An improved RT-PCR was developed and validated for the detection of Yam mild mosaic virus (YMMV). Sequences of the coat protein core region of 19 Chinese isolates were obtained, and analysis indicated the presence of different genetic variants.

Phylogenetic analyses showed that the Chinese isolates were divided into two distinct clusters. Complete genomic sequences of two distinct Chinese variants were determined to be 9527 and 9529 nucleotides long, excluding the 3′ poly (A) tail. Their genomic structure and organization were virtually identical to that of a Brazilian isolate. The two variants shared identity of 87.3% to one another and 83.9–84.6% to the Brazilian variant at the genomic sequence level. Phylogenetic analyses supported that they represented two distinct YMMV lineages. “
“The penetration process and defence reactions (hypersensitive response, oxidative burst and cell wall fortification) of Colletotrichum orbiculare were studied histochemically on pepper cultivar ‘A11’ (non-host) and susceptible Non-specific serine/threonine protein kinase cucumber cultivar ‘Changchun Thorn’ (host). The results indicate that C. orbiculare could hardly penetrate the non-host pepper leaves. It was papillae rather than hypersensitive response and H2O2 that played an important role in resisting the colonization and development of C. orbiculare on the non-host pepper. The depolymerization of the actin microfilament weakened the papilla deposition of pepper and allowed successful penetration of the non-adapted C. orbiculare, suggesting that the actin cytoskeleton of pepper is significant in preventing the invasion of the non-host pathogen C. orbiculare.

1%). Single and multiple infections were found in 42.38% and 16.54% of the samples, respectively. The most common multiple infection was of ToMV, PVY or both. These results show that the percentage of infected plants and plots in open field cultivation is very high in this region and the origin of the seed is an important factor in the incidence of virus selleck infection. In this respect, preventive measures, including virus-free

seed, resistant cultivars and improved cultural practices, could reduce the incidence of virus infection. “
“The distinguished plant cell wall component referred to as hydroxyproline-rich glycoproteins (HRGPs) exists in two forms: soluble in the symplast and insoluble in the apoplast. Insolubilization of HRGPs in cell walls through oxidative cross-linking which is elicited by stress represents a characteristic feature exhibited by two classes of HRGPs, namely, extensins and proline/HRGPs. Cross-linking of these HRGPs is an important process to strengthen the find more cell walls that contributes to plant defence reactions. In this review, the available information on these proteins is analysed with respect

to their roles in host-pathosystems and the various techniques applied for their characterization. Future prospects on strengthening of cell walls through gene regulation and transgenic approaches are also addressed. “
“An improved RT-PCR was developed and validated for the detection of Yam mild mosaic virus (YMMV). Sequences of the coat protein core region of 19 Chinese isolates were obtained, and analysis indicated the presence of different genetic variants.

Phylogenetic analyses showed that the Chinese isolates were divided into two distinct clusters. Complete genomic sequences of two distinct Chinese variants were determined to be 9527 and 9529 nucleotides long, excluding the 3′ poly (A) tail. Their genomic structure and organization were virtually identical to that of a Brazilian isolate. The two variants shared identity of 87.3% to one another and 83.9–84.6% to the Brazilian variant at the genomic sequence level. Phylogenetic analyses supported that they represented two distinct YMMV lineages. “
“The penetration process and defence reactions (hypersensitive response, oxidative burst and cell wall fortification) of Colletotrichum orbiculare were studied histochemically on pepper cultivar ‘A11’ (non-host) and susceptible Verteporfin concentration cucumber cultivar ‘Changchun Thorn’ (host). The results indicate that C. orbiculare could hardly penetrate the non-host pepper leaves. It was papillae rather than hypersensitive response and H2O2 that played an important role in resisting the colonization and development of C. orbiculare on the non-host pepper. The depolymerization of the actin microfilament weakened the papilla deposition of pepper and allowed successful penetration of the non-adapted C. orbiculare, suggesting that the actin cytoskeleton of pepper is significant in preventing the invasion of the non-host pathogen C. orbiculare.

Strains of normally bioluminescent species always contained lcf even when they were found not to produce light, thus demonstrating the utility of this methodology as a powerful tool for identifying bioluminescent species. Bioluminescence and lcf were confined to the

Gonyaulacales, Noctilucales, and Peridiniales. Considerable variation was observed among genera, or even species within some genera, that contained this gene. Partial sequences of lcf were obtained for the genera Ceratocorys, Ceratium, Fragilidium, and Protoperidinium as well as from previously untested species or gene regions of Alexandrium and Gonyaulax. The sequences revealed Ibrutinib datasheet high variation among gene copies that obscured the boundaries between species or even genera, some of which could be explained by the presence of two genetic variants within the same species of Alexandrium. Highly divergent sequences within Alexandrium and Ceratium show a more diverse composition of lcf than previously known. “
“Keeping sterile stocks or cultures of microalgae is fundamental to microalgae biotechnology as well as basic scientific research. However, contamination by bacteria and/or fungi in microalgae cultures or stocks is often a problem. Here, we have developed a strategy for

reducing or eliminating bacterial www.selleckchem.com/products/FK-506-(Tacrolimus).html and fungal contamination by using a cocktail of antibiotics. Chlamydomonas reinhardtii P. A. Dang., a widely used unicellular green alga, has been used as a testing organism. A combination of ampicillin, cefotaxime, and carbendazim removed or reduced contamination by three different bacteria and two different fungi tested. A step-by-step procedure is provided, which is simple, economical, and effective. “
“Coccoid green algae are among the most widespread photosynthesizers in freshwater, marine, and terrestrial environments (Krienitz and Bock 2012). These little “green balls”

occur virtually in every type of aquatic and terrestrial habitat where photosynthesis is possible, and their existence was already known by phycologists of the 19th century, who provided the first species descriptions (e.g., Nägeli 1849). Due to their small size, general invisibility to the unaided eye, and uniform appearance, for Liothyronine Sodium a long time these algae did not attract substantial interest from scientists. The scarcity of morphological characters useful for their differentiation led to the general impression that coccoid green algae represented a homogeneous, poorly diversified group of organisms. This is reflected in the history of their classification: until a few decades ago, green coccoids were united in the single-order Chlorococcales and subdivided into a relatively small number of genera. Ultrastructural and molecular data have now revolutionized this view. In the last 20 years, coccoid green algae have been an endless source of surprises, and still continue to be.

4 This is unlike treatment with directly targeting antivirals, such as those for HCV or HIV, where treatment failure is often associated with drug resistance that can affect responsiveness to subsequent courses of treatment. Other examples where bridging analyses have impacted regulatory decision making include oxcarbazepine, topiramate, clevidipine,

and levofloxacin, to name a few.5-8 Bridging knowledge to provide clinical evidence of effectiveness and to support dosing recommendations not only is acceptable from a regulatory perspective, but when scientifically supported and warranted it is also encouraged to increase the efficiency by which new drugs and optimal dosing recommendations are made available to patients in need.9 This report summarizes the rationale to support the BOC dosing recommendations in prior P/R-null responders and treatment-naïve BOC late responders.10 BOC, boceprevir; FDA, U.S. Food selleck inhibitor and Drug Administration; HCV, hepatitis C virus; P/R, peginterferon alpha and ribavirin; RGT, response-guided therapy; SVR, sustained virologic response. The SPRINT-II and RESPOND-II

trial designs are illustrated find more in Fig. 1. The primary endpoint for both SPRINT-II and RESPOND-II was the proportion of subjects with SVR (sustained virologic response), defined as HCV RNA undetected at 24 weeks after the last dose of the study drug. Both trials had three treatment arms: (1) P/R for 48 weeks (Arm 1, P/R); (2) a response-guided therapy (RGT) arm with P/R lead-in for 4 weeks, followed by P/R with BOC for 24 weeks in SPRINT-II and 32 weeks in RESPOND-II (Arm 2, BOC-RGT); Racecadotril and (3) P/R lead-in for 4 weeks, followed by BOC with P/R for 44 weeks (BOC44) (Arm 3, BOC44). In SPRINT-II subjects randomized to the RGT arm who had HCV RNA undetected at weeks 8 through week 24 stopped all treatment

at week 28 and were classified as BOC treatment-naïve early responders. The remaining subjects in the RGT arm, who had HCV RNA detected at treatment week 8 or any subsequent week, but who had HCV RNA undetected at week 24, stopped BOC at week 28 but received P/R through week 48. These subjects are referred to as BOC treatment-naïve late responders. In the RGT arm of RESPOND-II treatment-experienced early responders (subjects with HCV RNA undetected at weeks 8 through 12) stopped all treatment at week 36, whereas treatment-experienced late responders (HCV RNA detected at week 8 but HCV RNA undetected at week 12) stopped BOC at week 36 but continued P/R through week 48. Although prior P/R null responders were excluded from RESPOND-II, the sponsor proposed that data from treatment-naïve subjects could be used to estimate the treatment effect in prior null responders because included among treatment-naïve subjects are a subset of subjects who are intrinsically poor responders to interferon.

Egg mass, incubation length and hatching success (89%) were similar for the 28 and 28 ± 3°C groups, whereas the 28 ± 6°C group only had a 5% hatching success, and the incubation length was 10 days longer. Upon hatching, there was no significant difference in body mass or straight carapace length between the 28 and 28 ± 3°C groups, and within the first 8 weeks ABC294640 clinical trial of hatching, there was no significant difference in growth rate, self-righting

time, crawling speed and swimming performance. A single survivor from the 28 ± 6°C group had a body mass that was 27% less compared with the other two groups and it did considerably poorer in all the performance tests. The study findings illustrated that daily fluctuations in incubation temperature up to 6°C had no effect upon hatchling E. macrurus phenotype, but there was a limit (12°C) by which the extent and recurrence of these fluctuations became detrimental. These thermal regimes are not yet apparent in the wild but will occur within the

geographical range of this species according to climate change predictions. “
“Many mammal species reproduce seasonally because of annual fluctuations in temperature, rainfall and photoperiod in often nutritionally challenging habitats. The reproductive biology of many small southern African mammals is largely unknown and in critical need LGK-974 supplier of study. We investigated the breeding pattern of the female spiny mouse (Acomys spinosissimus) from South Africa. We examined the ovarian development, follicular growth, circulating plasma progesterone concentrations and the reproductive status of wild-caught adult female spiny mice sampled over a 12-month period while also correcting for body mass

and age. From these data, we conclude that female A. spinosissimus breed seasonally. The main breeding season of the spiny mouse is between September and January, with plasma progesterone concentrations being elevated, ovarian volume and primary, secondary, tertiary and Graafian follicle numbers as ADP ribosylation factor well as the corpora body number being the highest and pregnancies occurring during this period. Females were reproductively inactive from February through to August. The breeding season coincides with the onset of the rainy season in the habitat, which starts around September and ends in April. Rainfall, in association with an increase in primary productivity and hence higher food availability, might be the most important factor shaping reproduction in the female spiny mouse. “
“Biology and Environmental Science, University of Sussex, Sussex, Brighton, UK From insects to mammals, many animals engage in behaviours known to follow cyclic patterns over days (e.g. singing, diving or foraging behaviours). Many of them are regulated by external factors, such as light intensity, and are thus associated with sunrise, sunset or zenith.

For example, SHCC and SLHCC had longer survival time than NHCC and the medium overall cumulative survival of SHCC, SLHCC, and NHCC were 56.7, 33.3, and 15.3 months, respectively (P = 0.004; Fig. 1G). In line with this result, the median disease-free survival of SLHCC was 28.0 months,

which was significantly better than that of NHCC (14.0 months), but similar to that of SHCC (38.0 months, P = 0.003; Fig. 1H). These results indicated that miR-140-5p might play a critical Sunitinib in vivo role in HCC metastasis and progression. To determine the biological significance of miR-140-5p in HCC metastasis, we performed a wound-healing assay and transwell assay using HCCLM3 and MHCC97-H cells. It was noted that ectopic miR-140-5p expression significantly suppressed wound healing in the studies with both HCCLM3 and MHCC97-H cells (P < 0.01; Fig. 2A). Transwell assays with Matrigel further demonstrated that miR-140-5p markedly inhibited the invasive capacity of HCCLM3 and MHCC97-H cells as compared with that of vector-transduced control cells (P < 0.001; Fig. 2B). To further confirm these results, we next examined miR-140-5p-induced cytoskeletal and morphologic changes in HCCLM3 cells. As shown in Fig. ABT-263 in vitro 2C, miR-140-5p stimulated the reorganization of F-actin leading to the formation of stress fiber-like structures, while these structures were absent in vector-transduced cells. To demonstrate the effect of miR-140-5p on HCC growth, we performed an HCC cell proliferation

assay. As shown in crotamiton Fig. 2D, lentiviral-induced ectopic miR-140-5p resulted in a significant decrease in cell proliferation in both HCCLM3 and MHCC97-H cells (Fig.

2D). We then performed cell cycle analysis and revealed that miR-140-5p arrested the cells at S phase. Ectopic miR-140-5p expression decreased the percentage of cells in G1 phase from 60.92% to 38.64% (P < 0.001), but increased the percentage of cells in S phase and G2/M phase from 34.82% to 53.43% (P < 0.001) and 4.26% to 7.93% (P > 0.05), respectively (Fig. 2E). Consistent with these results, ectopic miR-140-5p expression suppressed colony formation (Fig. 2F). To confirm the above data in vivo, we did xenotransplantation of tumor grafts. Consistently, miR-140-5p significantly inhibited tumor growth in vivo. The size of subcutaneous tumors and local liver tumors originated from miR-140-5p-transduced HCCLM3 cells were dramatically smaller than that of vector-transduced cells (P = 0.022, P = 0.013, respectively; Fig. 3A). We further examined the mice for liver and lung metastasis of the carcinoma cells. As shown in Fig. 3B, the intrahepatic metastasis rates in mice with miR-140-5p-transduced grafts was only 20%, while metastasis was completely absent in the lung. In contrast, mice engrafted with vector-transduced tumors showed 80% and 60% rates for intrahepatic and lung metastasis, respectively. Taken together, our data support the conclusion that miR-140-5p suppresses HCC growth and metastasis.