Synthesis and biological evaluation of 1-phenyl-tetrahydro-ß-carboline-based first dual PRMT5/EGFR inhibitors as potential anticancer agents.

Protein arginine methyltransferase 5 (PRMT5) and epidermal growth factor receptor (EGFR) are both involved in the regulation of various cancer-related processes, and their dysregulation or overexpression has been observed in many types of tumors. In this study, we designed and synthesized a series of 1-phenyl-tetrahydro-ß-carboline (THßC) derivatives as the first class of dual PRMT5/EGFR inhibitors. Among the synthesized compounds, 10p showed the most potent dual PRMT5/EGFR inhibitory activity, with IC50 values of 15.47 ± 1.31 and 19.31 ± 2.14 µM, respectively. Compound 10p also exhibited promising antiproliferative activity against A549, MCF7, HeLa, and MDA-MB-231 cell lines, with IC50 values below 10 µM. Molecular docking studies suggested that 10p could bind to PRMT5 and EGFR through hydrophobic, π-π, and cation-π interactions. Furthermore, 10p displayed favorable pharmacokinetic properties and oral bioavailability (F = 30.6%) in rats, and administrated orally 10p could significantly inhibit the growth of MCF7 orthotopic xenograft tumors. These results indicate that compound 10p is a promising hit compound for the development of novel and effective dual PRMT5/EGFR inhibitors as potential anticancer agents.

Nickel-Catalyzed Unsymmetrical Bis-Allylation of Alkynes.

The catalytic bis-allylation of alkynes is an important but challenging protocol to construct all-carbon tetra-substituted alkenes. Particularly, the catalytic unsymmetrical bis-allylation of alkynes remains as an underexplored task to date. We herein report an unprecedented unsymmetrical bis-allylation by simultaneously utilizing electrophilic trifluoromethyl alkene and nucleophilic allylboronate as the allylic reagents. With the aid of robust Ni0 /NHC catalysis, valuable skipped trienes can be obtained in high regio- and stereo-selectivities under mild conditions. Mechanistic studies indicate that the reaction may proceed through a ß-fluorine elimination of a nickelacycle followed by a transmetalation step with allylboronate. The present method exhibits a good tolerance of various functional groups. Besides, the skipped triene products can undergo an array of elaborate transformations, which highlights the potential applications of this strategy.

PELO facilitates PLK1-induced the ubiquitination and degradation of Smad4 and promotes the progression of prostate cancer.

PLK1 and Smad4 are two important factors in prostate cancer initiation and progression. They have been reported to play the opposite role in Pten-deleted mice, one is an oncogene, the other is a tumor suppressor. Moreover, they could reversely regulate the PI3K/AKT/mTOR pathway and the activation of MYC. However, the connections between PLK1 and Smad4 have never been studied. Here, we showed that PLK1 could interact with Smad4 and promote the ubiquitination and degradation of Smad4 in PCa cells. PLK1 and PELO could bind to different domains of Smad4 and formed a protein complex. PELO facilitated the degradation of Smad4 through cooperating with PLK1, thereby resulting in proliferation and metastasis of prostate cancer cell. Changes in protein levels of Smad4 led to the alteration of biological function that caused by PLK1 in prostate cancer cells. Further studies showed that PELO upregulation was positively associated with high grade PCa and knockdown of PELO expression significantly decreased PCa cell proliferation and metastasis in vitro and vivo. PELO knockdown in PCa cells could enhance the tumor suppressive role of PLK1 inhibitor. In addition, blocking the interaction between PELO and Smad4 by using specific peptide could effectively inhibit PCa cell metastasis ability in vitro and vivo. Overall, these findings identified a novel regulatory relationship among PLK1, Smad4 and PELO, and provided a potential therapeutic strategy for advanced PCa therapy by co-targeting PLK1 and PELO.

Two Classes of Myosin Inhibitors, Para-nitroblebbistatin and Mavacamten, Stabilize ß-Cardiac Myosin in Different Structural and Functional States.

In addition to a conventional relaxed state, a fraction of myosins in the cardiac muscle exists in a low-energy consuming super-relaxed (SRX) state, which is kept as a reserve pool that may be engaged under sustained increased cardiac demand. The conventional relaxed and the super-relaxed states are widely assumed to correspond to a structure where myosin heads are in an open configuration, free to interact with actin, and a closed configuration, inhibiting binding to actin, respectively. Disruption of the myosin SRX population is an emerging model in different heart diseases, such as hypertrophic cardiomyopathy, which results in excessive muscle contraction, and stabilizing them using myosin inhibitors is budding as an attractive therapeutic strategy. Here we examined the structure-function relationships of two myosin ATPase inhibitors, mavacamten and para-nitroblebbistatin, and found that binding of mavacamten at a site different than para-nitroblebbistatin populates myosin into the SRX state. Para-nitroblebbistatin, binding to a distal pocket to the myosin lever arm near the nucleotide-binding site, does not affect the usual myosin SRX state but instead appears to render myosin into a new, perhaps much more inhibited, 'ultra-relaxed' state. X-ray scattering-based rigid body modeling shows that both mavacamten and para-nitroblebbistatin induce novel conformations in human ß-cardiac heavy meromyosin that diverge significantly from the hypothetical open and closed states, and furthermore, mavacamten treatment causes greater compaction than para-nitroblebbistatin. Taken together, we conclude that mavacamten and para-nitroblebbistatin stabilize myosin in different structural states, and such states may give rise to different functional energy-sparing states.

Loss of CDH1 promotes the metastasis of hypopharyngeal squamous cell carcinoma through the STAT3-MMP-9 signaling pathway.

BACKGROUND: Distant metastasis is the major cause of death in patients with hypopharyngeal squamous cell carcinoma (HSCC). CDH1 is correlated with tumor invasion and metastasis; however, its function in HSCC remains unclear. METHODS: We used immunohistochemistry (IHC) staining to evaluate the expression of CDH1 in 31 and 78 specimens from primary HSCC patients with and without postoperative lung metastases respectively. Sulforhodamine B (SRB) and CCK-8 assays were used to test the proliferation of HSCC cells. Motility of HSCC cells was investigated by migration and invasion assays. Western blot analysis was used to measure the levels of CDH1 and other proteins. RESULTS: We found that the low expression of CDH1 was significantly associated with postoperative lung metastasis in HSCC (P<0.001). Moreover, CDH1 was reduced concomitantly with the upregulation of MMP-9 in the same HSCC sample. Further mechanistic investigation showed that silencing CDH1 elevated the level of MMP-9, which was coupled with the phosphorylation of STAT3. Subsequently, inhibiting STAT3 either by siRNA transfection or by pharmacological suppression with AG490 attenuated MMP-9 upregulation and prevented the enhanced proliferation and invasion caused by CDH1 loss in FaDu cells. CONCLUSIONS: CDH1 plays vital roles in HSCC metastasis and might serve as a potential therapeutic target for the clinical treatment of HSCC.

Notch1 serves as a prognostic factor and regulates metastasis via regulating EGFR expression in hypopharyngeal squamous cell carcinoma.

OBJECTIVE: Hypopharyngeal squamous cell carcinoma (HSCC) remains one of the most lethal malignancies in head and neck. Notch1 has been validated to play prominent roles in the occurrence and development of various types of cancer. The aim of this study was to explore the function and underlying mechanism of Notch1 in HSCC. PATIENTS AND METHODS: Seventy-one cancer tissue samples and adjacent noncancerous formalin-fixed paraffin embedded tissue specimens were analyzed by immunohistochemistry. As Notch1 is overexpressed in HSCC, we further questioned whether there was a relationship between Notch1 and the clinicopathological characteristics. After confirming the successful knockdown of Notch1 by siRNA, the migration and invasion after gene knockdown were investigated by Transwell chambers. We then tried to identify YBX1 and EGFR expression using real-time PCR (RT-PCR) and Western blot analyses. To further determine whether the downexpression of EGFR was caused by YBX1 and the overexpression of YBX1 was caused by gene amplification, the expression of EGFR was detected by RT-PCR and Western blot assays. RESULTS: We found that the expression of Notch1 and EGFR in HSCC tissues was upregulated compared with those in the adjacent noncancerous tissues. Further clinicopathological characteristics analysis revealed that the expression of Notch1 was positively correlated with distant metastasis (P=0.003) and tumor differentiation (P=0.031). The high expression of Notch1 is an independent prognostic factor for a poor overall survival in patients with HSCC (P=0.015, χ 2=10.403). Knocking down of Notch1 significantly inhibits the migration and invasion of FaDu cells in vitro. Mechanistic investigation reveals that Notch1 knockdown is found suppressing the expression of EGFR at transcriptional level. Interestingly, we further found that Notch1 knockdown also decreased the expression of YBX1, which is a transcription factor of EGFR. Moreover, the upregulation of YBX1 reverses the suppression of Notch1 on EGFR. Furthermore, forced overexpression of YBX1 induced the invasion of FaDu cells. CONCLUSION: Taken together, we found a positively cross-linked role of Notch1 signaling in the outcome of HSCC, providing a novel valuable prognostic marker and potential therapeutic target for the treatment of HSCC patients. Notch1 is a core signaling molecule for regulating migration and invasion via interplaying with EGFR in HSCC cells.

Characterization of a non-nudix pyrophosphatase points to interplay between flavin and NAD(H) homeostasis in Saccharomyces cerevisiae.

The flavin cofactors FMN and FAD are required for a wide variety of biological processes, however, little is known about their metabolism. Here, we report the cloning and biochemical characterization of the Saccharomyces cerevisiae pyrophosphatase Fpy1p. Genetic and functional studies suggest that Fpy1p may play a key role in flavin metabolism and is the first-reported non-Nudix superfamily enzyme to display FAD pyrophosphatase activity. Characterization of mutant yeast strains found that deletion of fpy1 counteracts the adverse effects that are caused by deletion of flx1, a known mitochondrial FAD transporter. We show that Fpy1p is capable of hydrolyzing FAD, NAD(H), and ADP-ribose. The enzymatic activity of Fpy1p is dependent upon the presence of K+ and divalent metal cations, with similar kinetic parameters to those that have been reported for Nudix FAD pyrophosphatases. In addition, we report that the deletion of fpy1 intensifies the FMN-dependence of null mutants of the riboflavin kinase Fmn1p, demonstrate that fpy1 mutation abolishes the decreased fitness resulting from the deletion of the flx1 ORF, and offer a possible mechanism for the genetic interplay between fpy1, flx1 and fmn1.

Hypomorphic Recessive Variants in SUFU Impair the Sonic Hedgehog Pathway and Cause Joubert Syndrome with Cranio-facial and Skeletal Defects.

The Sonic Hedgehog (SHH) pathway is a key signaling pathway orchestrating embryonic development, mainly of the CNS and limbs. In vertebrates, SHH signaling is mediated by the primary cilium, and genetic defects affecting either SHH pathway members or ciliary proteins cause a spectrum of developmental disorders. SUFU is the main negative regulator of the SHH pathway and is essential during development. Indeed, Sufu knock-out is lethal in mice, and recessive pathogenic variants of this gene have never been reported in humans. Through whole-exome sequencing in subjects with Joubert syndrome, we identified four children from two unrelated families carrying homozygous missense variants in SUFU. The children presented congenital ataxia and cerebellar vermis hypoplasia with elongated superior cerebellar peduncles (mild "molar tooth sign"), typical cranio-facial dysmorphisms (hypertelorism, depressed nasal bridge, frontal bossing), and postaxial polydactyly. Two siblings also showed polymicrogyria. Molecular dynamics simulation predicted random movements of the mutated residues, with loss of the native enveloping movement of the binding site around its ligand GLI3. Functional studies on cellular models and fibroblasts showed that both variants significantly reduced SUFU stability and its capacity to bind GLI3 and promote its cleavage into the repressor form GLI3R. In turn, this impaired SUFU-mediated repression of the SHH pathway, as shown by altered expression levels of several target genes. We demonstrate that germline hypomorphic variants of SUFU cause deregulation of SHH signaling, resulting in recessive developmental defects of the CNS and limbs which share features with both SHH-related disorders and ciliopathies.

Elevated AEG-1 expression in macrophages promotes hypopharyngeal cancer invasion through the STAT3-MMP-9 signaling pathway.

Macrophages play a critical role in tumor invasion and metastasis, which remain major causes of mortality in patients with hypopharyngeal cancer. Here we investigate the effect of an oncogene, AEG-1 expressed in macrophages on the invasion of hypopharyngeal cancer cells. AEG-1 is more highly expressed in macrophages of human hypopharyngeal cancer samples compared with adjacent non-tumor controls. Using matrigel invasion assay system, THP-1-derived macrophages with forced AEG-1 overexpression enhance FaDu cell invasion whereas macrophages with AEG-1 silence inhibit. Matrix metalloproteinase 9 (MMP-9), which is important in tumor invasion and metastasis through degrading extracellular matrix, is up-reulated by AEG-1 partly through NF-κB p65 in macrophages. Intriguingly, macrophage AEG-1 also induces MMP-9 up-regulated expression in FaDu cells. Furthermore, macrophage AEG-1 activates signal transducer and activator of transcription 3 (STAT3) in FaDu cells, which is responsible for macrophage AEG-1-induced an increase in MMP-9 expression and invasion of FaDu cells. This is the first to demonstrate that macrophage AEG-1 promotes tumor invasion through up-regulation of MMP-9 in both macrophages and cancer cells. Thus, the results provide evidences that macrophage AEG-1 contributes to promotion of tumor invasion, and represents as a potential target in hypopharyngeal cancer therapy.

Afatinib down-regulates MCL-1 expression through the PERK-eIF2α-ATF4 axis and leads to apoptosis in head and neck squamous cell carcinoma.

Afatinib is the second generation of irreversible inhibitor of EGFR, HER2 and HER4, which has shown encouraging phase II and III clinical outcomes in the treatment of head and neck squamous cell carcinoma (HNSCC). However, the molecular mechanism of afatinib-induced apoptosis in HNSCC is poorly understood. In the present investigation, we discovered that down-regulation of MCL-1, an anti-apoptotic member of BCL-2 family, was responsible for afatinib-triggered apoptosis. And the inactivation of AKT-mTOR signaling caused by afatinib lead to translational inhibition of MCL-1 expression. As a crucial branch of ER stress, PERK-eIF2α-ATF4 axis was also stimulated in HNSCC cells after afatinib incubation. Silencing either eIF2α or ATF4 by siRNA transfection relieved afatinib-caused suppression of AKT-mTOR activity, attenuating MCL-1 down-regulation as well as subsequent apoptosis. Collectively, the results show that afatinib hampers AKT-mTOR activation by stimulating PERK-eIF2α-ATF4 signaling pathway, giving rise to MCL-1 down-regulation mediated apoptosis in HNSCC cells. Therefore, our findings reveal the elaborate molecular network of afatinib-induced apoptosis in HNSCC, which would provide substantial theoretical underpinnings for afatinib clinical application and highlight its promising prospect in HNSCC treatment.

Two New Anthraquinones from the Roots of Prismatomeris connata.

Two new anthraquinones, 4-hydroxy-1,2,3-trimethoxy-7-hydroxymethylanthracene-9,10-dione (1) and 1,2,3-trimethoxy-7-hydroxymethylanthracene-9,10- dione (2), were isolated from the roots of Prismatomeris connata, a Chinese medicinal herb. Their structures were elucidated by spectroscopic analysis. Compound 1 exhibited cytotoxicity against a panel of H1229, HTB 179, A549 and H520 lung tumor cell lines with IC50 values ranging from 12.3 to 20 µM.

[miR-140-5p affects the migration and invasion of hypopharyngeal carcinoma cells by downregulating ADAM10 expression].

OBJECTIVE: To investigate the expression of miR-140-5p and ADAM10 in hypopharyngeal carcinoma tissues and their effects on the migration and invasion of FaDu cells and underlying mechanism. METHODS: The miR-140-5p and ADAM10 mRNA levels in 33 cases of hypopharyngeal carcinoma tissues and adjacent normal tissues were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Transwell migration assay and transwell invasion assay were used to test the metastasis ability of FaDu cells after upregulation or downregulation of miR-140-5p and downregulation of ADAM10. The protein expression levels of ADAM10 in hypopharyngeal carcinoma tissues and the FaDu cells after transfection were determined by Western blot assays. RESULTS: The expression level of miR-140-5p was significantly downregulated in hypopharyngeal carcinoma tissues compared with adjacent tissues (t=-4.016, P<0.01), which was significantly correlated with tumor classification and lymph node metastasis (P<0.05). Conversely, mRNA and protein expressions of ADAM10 were significantly upregulated in tumor tissues (t=3.960, P<0.01; t=12.089, P<0.01), and were significantly downregulated in the FaDu cells after tranfected with si-ADAM10 (t=8.653, P<0.05; t=5.191, P<0.05). Transwell assay showed that compare with control group, the migration and invasive cells decreased significantly in hsa-mir-140-5p group (t=3.255, P<0.05; t=2.942, P<0.05), while increased significantly in anti-hsa-mir-140-5p group, (t=-13.521, P<0.05; t=-6.223, P<0.05). The migration and invasive cells in si-ADAM10 group were less than those in control group (t=4.759, P<0.05; t=3.663, P<0.05). The downregulation of ADAM10 attenuated the effect of anti-mir-140-5p in FaDu cells. Western blot assay showed that ADAM10 expression was apparently decreased in hsa-mir-140-5p group and increased in anti-mir-140-5p group compared with control group. CONCLUSIONS: The expression of miR-140-5p was significantly downregulated in hypopharyngeal carcinoma tissues and correlated with tumor classification and lymph node metastasis. ADAM10 was upregulated in tumor tissues. MiR-140-5p suppresses the migration and invasion abilities of FaDu cells, possibly through downregulation of ADAM10.

MicroR-140-5p suppresses tumor cell migration and invasion by targeting ADAM10-mediated Notch1 signaling pathway in hypopharyngeal squamous cell carcinoma.

MicroRNAs (miRNAs) are small non-coding RNAs of approximately 22 nucleotides that negatively regulate gene expression at the post-transcriptional level. Downexpression of miR-140-5p was reported in some human cancers, and combined with a reduction of cell migration and invasion, suggesting that miR-140-5p functions as a tumor suppressor. However, little is known about the expression and function of miR-140-5p in hypopharyngeal squamous cell carcinoma (HSCC). In this research, we found that miR-140-5p was significantly downregulated in HSCC tissues and correlated to tumor classification and lymph node metastasis. Restoration of miR-140-5p suppressed the migration and invasion of FaDu cells, and decreased the protein expression levels of ADAM10. Furthermore, the luciferase reporter assay revealed that miR-140-5p was directly bound to ADAM10 mRNA and knockdown of ADAM10 could inhibit FaDu cell migration and invasion and reduced the protein expression levels of and Notch1 intracellular domain (NICD1). Of note, knockdown of Notch1 could inhibit the migration and invasion of FaDu cells and rescued the effect of miR-140-5p inhibitor in FaDu cells. Taken together, our study demonstrates that miR-140-5p suppresses tumor migration and invasion by inhibiting ADAM10-mediated Notch1 signaling pathway and suggests that miR-140-5p could have potential therapeutic applications in HSCC.

Epithelial cell adhesion molecule is overexpressed in hypopharyngeal carcinoma and suppresses the metastasis and proliferation of the disease when downregulated.

The epithelial cell adhesion molecule (EpCAM) is overexpressed in the majority of human epithelial carcinomas, and its overexpression is associated with proliferation and neoplastic transformation. However, the precise molecular mechanism involved in EpCAM-related proliferation and metastasis in hypopharyngeal carcinoma is unknown. The aim of the present study was to identify the role of EpCAM in the metastasis and proliferation of hypopharyngeal carcinoma. An immunohistochemical staining assay indicated that EpCAM was overexpressed in primary hypopharyngeal carcinoma tissues, and that this overexpression correlated with the tumor size and lymph node metastasis. In the following treatment of the hypopharyngeal carcinoma FaDu cell line with EpCAM, the downregulation of EpCAM was found to significantly suppress cell metastasis and proliferation, as detected by Transwell, clone formation and MTT assays. Additionally, western blot analysis revealed that EpCAM downregulation increased the expression of the adhesion- and proliferation-related factors, E-cadherin, α-catenin and ß-catenin, in the cytoskeleton, as well as ß-catenin expression in the nucleus. In conclusion, the present study indicated that EpCAM is a potential oncogene and contributes to the metastasis of hypopharyngeal carcinoma. The current study is the first to provide evidence for the potential value of targeting EpCAM in hypopharyngeal carcinoma therapy.

Tumor necrosis factor α induces epithelial-mesenchymal transition and promotes metastasis via NF-κB signaling pathway-mediated TWIST expression in hypopharyngeal cancer.

Epithelial-mesenchymal transition (EMT) is an important mechanism in cancer metastasis. Tumor necrosis factor α (TNFα) can induce cancer invasion and metastasis associated with EMT. However, the underlying mechanisms are not entirely clear. Therefore, we investigated whether TNFα has an effect on EMT and invasion and metastasis in human hypopharyngeal cancer FaDu cells, and further explored the potential mechanisms. In the present study, we demonstrated that TNFα induced EMT in FaDu cells and promoted FaDu cell migration and invasion. TNFα-induced EMT was characterized by a change from well organized cell-cell adhesion and cell polarity to loss of cell-cell contacts, cell scattering and increased expression of vimentin and N-cadherin accompanied by a decrease in E-cadherin. Furthermore, we found that p65 translocated to the nucleus after TNFα stimulation and increased the nuclear expression of TWIST. We demonstrated that TNFα treatment also increased the expression of TWIST by activating the NF-κB signaling pathway. While p65 was inhibited by siRNA-65 or BAY11-7082 (inhibitor of NF-κB), TWIST expression was also decreased. Therefore, we conclude that TNFα induces EMT and promotes metastasis via NF-κB signaling pathway-mediated TWIST expression in hypopharyngeal cancer.

[Expression and effect of microRNA-214 in advanced hypopharyngeal carcinoma].

OBJECTIVE: To investigate the expression of microRNA-214(miR-214) in advanced hypopharyngeal carcinoma tissues and its effects on the invasion, migration and colone formation of FaDu cells. METHODS: miR-214 expression in 30 cases of advanced hypopharyngeal carcinoma tissues and normal hypopharyngeal mucosa tissues was detected by real-time quantitative polymerase chain reaction (RT-qPCR). miR-214 was upregulated through transfecting the overexpression vector hsa-mir-214 into FaDu cells. The influences of miR-214 upregulation on the invasion, migration, clone formation and Twist expression were measured by Transwell invasion, Transwell migration, plate clone formation and Western blot assays, respectively. RESULTS: The expression of miR-214 in advanced hypopharyngeal carcinoma tissues (0.311 ± 0.206) was significantly less than normal hypopharyngeal mucosa tissues (1.620 ± 1.394; t = 5.09, P < 0.05) . The expression of miR-214 was notably upregulated after tranfected with hsa-mir-214 compared with the negative control group (t = 6.347, P < 0.05). The migration and invasion ability of FaDu cells transfeced with hsa-mir-214 was decreased by comparison with negative control cells (t = 11.6, P < 0.01; t = 6.499, P < 0.05). There was no significant difference of the average clony number and the cloning efficiency between the experimental and negative control groups (t = 0.592, P > 0.05). RESULTS: of Western blot assay showed that, Twist expression in the miR-214-overexpressed group was apparently decreased compared with that in the control group (t = 6.545, P < 0.05). CONCLUSIONS: miR-214 is expressed at a low level in advanced hypopharyngeal carcinoma tissues, and can obviously inhibit the invasion and migration abilities of FaDu cells, possibly because of its inhibiting effect on Twist expression. Additionally, miR-214 plays no significant role in the proliferation of FaDu cells.

[Effect of epithelial cell adhesion molecule on metastasis in hypopharyngeal carcinoma FaDu cells].

OBJECTIVE: To investigate the effect of knockdown of EpCAM by siRNA on invasion, migration, and colony abilities in hypopharyngeal carcinoma FaDu cells. METHODS: A siRNA against EpCAM was employed to inhibit the expression of EpCAM in FaDu cells. Measurements included the Transwell assay for invasion and migration, plate colony formation assay for cell colony ability, Western blot assay for EpCAM, E-cadherin, and ß-catenin expressions in total protein, cytoplasm, and cytoskeleton, respectively. RESULTS: mRNA and protein expressions of EpCAM were suppressed significantly in FaDu cells transfected by EpCAM siRNA (t = 6.46, P < 0.05; t = 10.25, P < 0.05) . Transwell assay showed in transwell assay, the average invasive cells in EpCAM siRNA cells (26.33 ± 3.71) was less than that in FaDu cells (61.47 ± 6.70; t = 7.95, P < 0.05)and control cells (54.13 ± 6.51; t = 6.42, P < 0.05); the average number of migration cells in EpCAM siRNA cells (79.87 ± 8.44) was lower than that in FaDu (167.53 ± 11.49; t = 10.90, P < 0.05) cells and control cells (162.13 ± 13.45; t = 8.97, P < 0.05). In plate colony formation assay, the average colony number of EpCAM siRNA cells was (78.00 ± 5.57), which was less than that of FaDu cells(177.30 ± 16.50; t = 9.78, P < 0.05) and control cells (173.67 ± 13.50; t = 11.35, P < 0.05). Western blot assays showed, silencing of EpCAM increased the expressions of E-cadherin (t = 4.58, P = 0.01) and ß-catenin (t = 3.76, P = 0.02) in cytoskeleton, and decreased the expressions of E-cadherin (t = 6.60, P < 0.05) and ß-catenin (t = 8.20, P < 0.05) in cytoplasm. CONCLUSIONS: The knockdown of EpCAM inhibits the invasion, migration, and colony formation abilities of FaDu cells, which is probably related to the regulation of E-cadherin and ß-catenin in cytoplasm and cytoskeleton, and EpCAM may be a promising gene therapy target for hypopharyngeal carcinoma.

Characteristics of hippocampal volumes in healthy Chinese from MRI.

BACKGROUND/OBJECTIVES: The volume of hippocampus has been associated with a number of disorders and would be helpful in clinical and research practice. This study provided the normal data of hippocampal volumes in a healthy Chinese sample and assessed the possible effects of age, gender and side on it. METHODS: Sixty-one normal participants with a broad age span from 6 to 82 years were enrolled in the study. The hippocampal volumes of them were obtained from oblique coronal MR images using inversion recovery (IR) sequence and normalized to adjust intersubject variation in head size. RESULTS: The corrected volume in the right side of hippocampus was 2.204-2.944 cm(3), and 2.068-2.700 cm(3) in the left. There were no statistically significant differences among different age and gender groups (p>0.05). The volume of right hippocampus was larger than that of the left side (p<0.001). CONCLUSIONS: We presented the volume range of hippocampus in healthy Chinese on MRI in this study. No age and gender effects were found on hippocampal volumes. Side-to-side asymmetry was obtained by statistical analysis.