HOXA5 inhibits the proliferation and metastasis of cervical squamous cell carcinoma by suppressing the ß-catenin/Snail signaling.

HOXA5, as a transcription factor, plays an important role in a variety of malignant tumors. Nevertheless, its biological role in cervical squamous cell carcinoma (CSCC) is largely unknown. In our study, we aimed to explore the function of HOXA5 in CSCC and its molecular mechanism. Immunohistochemistry showed that HOXA5 expression was downregulated in human CSCC tissues and HOXA5 staining was negatively correlated with tumor size and histological grade of CSCC. Ectopic expression of HOXA5 inhibited proliferative and metastatic abilities of CSCC cells in vitro and in vivo. Furthermore, overexpression of HOXA5 inhibited the cell cycle by arresting the S/G2 phase by flow cytometry and that was related to the downregulation of Cyclin A. Further study showed that HOXA5 suppressed EMT by inhibiting the ß-catenin/Snail signaling resulting in reduced metastasis of CSCC cells. Altogether, our results suggested that HOXA5 inhibited the proliferation and metastasis via repression of the ß-catenin/Snail pathway, proposing the potential role of HOXA5 in the prevention and treatment of CSCC.

Striatal D1 Dopamine Neuronal Population Dynamics in a Rat Model of Levodopa-Induced Dyskinesia.

BACKGROUND: The pathophysiology of levodopa-induced dyskinesia (LID) in Parkinson's disease (PD) is not well understood. Experimental data from numerous investigations support the idea that aberrant activity of D1 dopamine receptor-positive medium spiny neurons in the striatal direct pathway is associated with LID. However, a direct link between the real-time activity of these striatal neurons and dyskinetic symptoms remains to be established. METHODS: We examined the effect of acute levodopa treatment on striatal c-Fos expression in LID using D1-Cre PD rats with dyskinetic symptoms induced by chronic levodopa administration. We studied the real-time dynamics of striatal D1 + neurons during dyskinetic behavior using GCaMP6-based in vivo fiber photometry. We also examined the effects of striatal D1 + neuronal deactivation on dyskinesia in LID rats using optogenetics and chemogenetic methods. RESULTS: Striatal D1 + neurons in LID rats showed increased expression of c-Fos, a widely used marker for neuronal activation, following levodopa injection. Fiber photometry revealed synchronized overactivity of striatal D1 + neurons during dyskinetic behavior in LID rats following levodopa administration. Consistent with these observations, optogenetic deactivation of striatal D1 + neurons was sufficient to inhibit most of the dyskinetic behaviors of LID animals. Moreover, chemogenetic inhibition of striatal D1 + neurons delayed the onset of dyskinetic behavior after levodopa administration. CONCLUSION: Our data demonstrated that aberrant activity of striatal D1 + neuronal population was causally linked with real-time dyskinetic symptoms in LID rats.

Inhibition of striatal dopamine D5 receptor attenuates levodopa-induced dyskinesia in a rat model of Parkinson's disease.

Levodopa-induced dyskinesia (LID) is experienced by most patients of Parkinson's disease (PD) upon the long-term use of the dopamine precursor levodopa. Striatal dopaminergic signaling plays a critical role in the pathogenesis of LID through its interactions with dopamine receptors. The specific roles of striatal dopaminergic D5 receptors in the pathophysiological process of LID are still poorly established. In the study, we investigated the role of striatal dopamine D5 receptor in LID by using PD rats with or without dyskinetic symptoms after chronic levodopa administration. The experimental results showed that the expression level of D5 receptors in the sensorimotor striatum of dyskinetic rats is significantly higher than that of the non-dyskinetic controls. The administration of levodopa increased c-Fos expression in a subpopulation of sensorimotor striatum neurons of dyskinetic rats, but not in non-dyskinetic rats. The majority of the c-Fos+ neurons activated by levodopa in the striatum are positive for D5 receptor staining. Intrastriatal injection of D1-like (D1 and D5) dopamine receptor antagonist, SCH-23390, significantly inhibited dyskinetic behavior in dyskinetic rats after the injection of levodopa, meanwhile, intrastriatal administration of SKF-83959, a partial D5 receptor agonist, yielded significant dyskinetic movements in dyskinetic rats without levodopa. In contrast, intrastriatal perfusion of small interfering RNA directed against DRD5 downregulated D5 receptors expression and moderately inhibited dyskinetic behavior of dyskinetic animals. Our data suggested that the striatal dopamine D5 receptor might play a novel role in the pathophysiology of LID.

HNF1A/CASC2 regulates pancreatic cancer cell proliferation through PTEN/Akt signaling.

Pancreatic cancer (PC) has a high mortality rate in all cancers worldwide. According to recent studies, long noncoding RNA-CASC2 is involved in the development and progression of many malignant tumors; in the present study, we demonstrated that lncRNA-CASC2 was specifically downregulated in PC tissues and cell lines, and a lower CASC2 expression in PC was related with a poorer prognosis. CASC2 suppressed PC cell proliferation. Hepatocyte nuclear factor 1 alpha (HNF1A) is a transcription factor known to regulate pancreatic differentiation and maintain the homeostasis of the endocrine pancreas. Recently, HNF1A is considered to be a possible tumor suppressor in PC. In the present study, we observed that HNF1A positively regulated CASC2 expression. Through luciferase assays, we demonstrated that CASC2 gene possessed an HNF1A-responsive element (CASC2-HNF1A RE); HNF1A could promote CASC2 expression through direct binding to CASC2-HNF1A RE. Further, PTEN/Akt signaling was involved in HNF1A regulation of CASC2. Finally, we evaluated the expression level of HNF1A in PC tissues; lower HNF1A expression was correlated with shorter overall survival in patients with PC. Taken together, these findings will shed light on the role and mechanism of HNF1A/CASC2 in regulating PC cells proliferation through PTEN/Akt signaling. CASC2 may serve as a potential therapeutic target in PC in the future.

A Pilot Study of RSK4 Expression in Patients with Human Non-Small Cell Lung Carcinoma.

BACKGROUND: Lung cancer represents the leading cause of cancer deaths worldwide, with non-small cell lung carcinoma (NSCLC) comprising the most common type of lung cancer. OBJECTIVE: The aim of this pilot study was to ascertain RSK4 expression at both the gene and protein level in normal and cancerous tissue of patients with NSCLC. METHODS: From January 2015 to December 2015, a total of forty patients diagnosed with NSCLC who underwent surgery were recruited into this study. All NSCLC diagnoses were confirmed by pathological examination. Normal and cancerous lung tissues were collected via surgical dissection. RSK4 gene and protein expression levels were ascertained using reverse transcription-polymerase chain reaction amplification and immunohistochemistry, respectively. Expression levels and patient pathological clinical parameters were compared by semi-quantitative method. RESULTS: Both RSK4 gene and protein expression levels were significantly reduced in patient cancerous tissue compared to their normal lung tissue (P<0.01). CONCLUSIONS: RSK4 expression is associated with clinical and pathological staging of NSCLC. Our preliminary data from this pilot study suggest that RSK4 constitutes a putative tumor suppressor gene for NSCLC. The relationship between RSK4 gene expression and NSCLC prognosis and clinical outcome needs to be further studied in a multi-center cohort study with a large patient sample.

Expression and clinical significance of HOXA5, E-cadherin, and ß-catenin in cervical squamous cell carcinoma.

OBJECTIVES: HOXA5 has been identified as a biomarker in pathogenesis of several cancers, such as non-small cell lung cancer (NSCLC) and breast cancer cells. The role has not been explored in cervical squamous cell carcinoma (CSCC). METHODS: Tissues of 120 cases with CSCC and 30 controls with chronic cervicitis were constructed from our archived surgical pathology files and staining with HOXA5. Additional antibodies to E-cadherin and ß-catenin were stained for comparison. For each marker, low expression was defined as staining score 0 to 3 points, whereas high expression referred to 4 points and above. Fifty-four patients in this research with cervical cancer were followed up for prognostic assessment. RESULT: HOXA5 had high expression in chronic cervicitis and low in CSCC (P=0.004). The positivity rates of HOXA5 in patients without muscular layer invasion (MLI) and lymphatic invasion (LI) was higher than that in metastasis (113 vs. 17; 117 vs. 3). Consistently, low expression of HOXA5 was more common in poorly differentiated carcinoma, CSCC subjects without MLI and LI. Expression of E-cadherin and ß-catenin was parallel with the expression of HOXA5. Additionally, patients with higher expression of HOXA5 had much more favorable prognosis than those with lower expression among follow up of the 54 patients. CONCLUSION: In parallel with E-cadherin and ß-catenin, low expression of HOXA5 was more common in CSCC patients with poor differentiation, without MLI and LI, among those which showed poor prognosis.

Prognostic value of phosphorylated Raf kinase inhibitory protein at serine 153 and its predictive effect on the clinical response to radiotherapy in nasopharyngeal carcinoma.

BACKGROUND: Radiation is an effective treatment against nasopharyngeal carcinoma (NPC). However, radioresistance-induced locoregional recurrence remains as a major cause of treatment failure. Therefore, radiosensitivity indicators prior to treatment should be developed to screen radioresistant patients. Previous studies revealed that RKIP (Raf kinase inhibitor protein) is associated with NPC prognosis and radiosensitivity. However, the relationship of p-Ser153 RKIP (RKIP in a phosphorylated form at residue serine153) expression with the effect of radiation and prognosis of NPC patients is not elucidated. Thus, these clinical implication of the phosphorylated RKIP in NPC has yet to be described. METHODS: The effect of p-Ser153 RKIP on locoregional relapse-free survival (LRRFS) was first analyzed in a retrospective cohort of NPC patients without distant metastasis at initial diagnosis. They received radical intensity-modulated radiotherapy alone. Of 180 patients were enrolled in the ongoing matched pair study. The patients were re-classified into radioresistant group or radiosensitive group on the basis of the specified criteria. Patients in the two groups were matched in terms of radiosensitivity-related factors. p-Ser153 RKIP was examined by immunohistochemical staining on a NPC tissue microarray before radiotherapy. The relationship between the expression of p-Ser153 RKIP and the effect of radiotherapy was also analyzed. RESULTS: In this study, a retrospective cohort with 733 cases who received radical radiotherapy alone was established. Using the cohort, we validated that the p-Ser153 RKIP expression observed through immunohistochemical staining in a pretreatment NPC tissue microarray was an independent prognostic factor of LRRFS and OS; we also confirmed that endemic patients with a positive p-Ser153 RKIP expression benefited from irradiation alone in terms of locoregional relapse-free survival. A total of 180 patients were enrolled in a matched pair study. Both groups were well matched in terms of radiosensitivity-related factors. On the basis of the p-Ser153 RKIP expression, we predicted the following data: 80.0 % sensitivity, 73.3 % specificity, 76.7 % accuracy, 75.0 % positive predictive value, and 78.6 % negative predictive value. CONCLUSIONS: Our results revealed for the first time that positive p-Ser153 RKIP expression was a favorable prognostic factor. It was also positively correlated with the radiosensitivity of NPC. p-Ser153 RKIP could also be used as a biomolecular marker with good availability and authenticity to preliminarily screen NPC-related clinical radiosensitivity.

The clinical value of LRRC3B gene expression and promoter hypermethylation in breast carcinomas.

Epigenetic alteration of tumor suppressor genes by promoter hypermethylation has played a key role in tumorigenesis, which is an important mechanism as indispensable as gene deletion and mutation. LRRC3B is a potential tumor suppressor gene newly discovered; however, the specific biologic function is still unknown. In the present study, we tested the expression levels of LRRC3B by methods associated with immunohistochemistry, Real-Time PCR, and methylation-specific polymerase chain reaction. Results showed that the expression levels were significantly low irrespective of methylation status, suggesting that there were other factors involved in this process. However, the expression profile of LRRC3B had a significant relationship with tissue grade, irrespective of the expressions of PR, CERB-2, VEGF, and Ki67 except in cases of p53 and ER, leading us to a conclusion that the abnormal expression of LRRC3B could serve as a useful marker for diagnosis and prognosis in breast carcinomas.

[Study on genetic aberrations of ocular mucosa-associated lymphoid tissue lymphomas occurring in southern China].

OBJECTIVE: To study the genetic aberrations of ocular extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type occurring in patients from southern China. METHODS: Fifty seven paraffin-embedded ocular MALT lymphoma specimens from patients in southern China were studied by interphase fluorescence-in-situ hybridization (FISH) for genetic aberrations including t(11;18)(q21;q21)/API2-MALT1, t(1;14)(p22;q32)/IgH-bcl-10, t(14;18) (q32;q21)/IgH-MALT1 and bcl-6/FOXP1 gene translocations. RESULTS: Amongst the 57 cases studied, 9 cases (15.8%) showed chromosome translocations, including 4 cases (7.0%) of t(11;18)(q21;q21)/API2-MALT1, 1 case (1.8%) of t(14;18) (q32;q21)/IgH-MALT1, 1 case (1.8%) of bcl-6 gene-related chromosome translocation and 3 cases (5.3%) of IgH-unknown translocation partner. FISH revealed 17 cases (29.8%) with 3 copies of bcl-6 gene, 21 cases (36.8%) with 3 copies of MALT1 gene and 12 cases (21.1%) with 3 copies of both genes. CONCLUSIONS: The MALT lymphoma-associated chromosome translocations t(11;18)(q21;q21)/API2-MALT1 and t(14;18) (q32;q21)/IgH-MALT1 are demonstrated in ocular MALT lymphomas of southern Chinese patients. The prevalence is significantly different from that reported in northern Chinese and northern American patients, indicating a geographic heterogeneity in the MALT lymphoma-associated genetic aberrations. The presence of 3 copies of bcl-6 and MALT1 genes is the commonest genetic abnormalities observed in ocular MALT lymphomas, suggesting a possible role in MALT lymphomagenesis.

[A clinical study of chromosome translocations in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue in Chinese patients].

OBJECTIVE: To investigate the genetic aberrations in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) lymphomas from different sites of the body in Chinese patients. METHODS: Two hundred and seventeen paraffin-embedded MALT lymphoma specimens from 11 major sites were studied with interphase fluorescence in situ hybridization (FISH) to detect t (11; 18) (q21; q21)/API2-MALT1, t (1; 14) (p22; q32)/IGH-BCL10, (14; 18) (q32; q21)/IGH-MALT1 and BCL6 gene involved chromosome translocations. RESULTS: These translocations were mutually exclusive and detected in 21% (46/217) of the cases, including t (11; 18) (q21; q21) API2-MALT1 13% (29/217), t (1; 14) (p22; q32) IGH-BCL10 in 1% (3/217), t (14; 18) (q32; q21) IGH-MALT1 1% (2/217), BCL6 involved translocation in 2% (4/217) and IGH-unknown translocation partner in 4% (8/217). t (11; 18) (q21; q21) API2-MALT1 was found with the highest frequency in MALT lymphoma from lungs (47%, 8/17) and small intestine (29%, 4/14), followed by salivary gland (17%, 1/6), stomach (14%, 12/84) and ocular adnexae (6%, 4/68). t (1; 14) (p22; q32) was only detected in lungs (12%, 2/17) and stomach (1%, 1/84). t (14; 18) (q32; q21) was mainly detected in lungs (6%, 1/17) and ocular adnexae (2%, 1/68). BCL6 gene involved translocation was detected in salivary gland (17%, 1/6) and stomach (4%, 3/84). CONCLUSIONS: It is demonstrated that the four translocations occur with markedly variable frequencies in MALT lymphoma of different sites in Chinese patients. The distributions of these chromosome translocations in Chinese patients are slightly different from those reported in western patients.

[Effects of angrographolide on plasma glucose level of experiment diabetic rats and its molecular mechanism].

OBJECTIVE: To investigate the effects of angrographolide on plasma glucose level of diabetic rats and its molecular mechanism. METHODS: The diabetic model rats were established by intraperitoneal injection of streptozotocin (65 mg/kg). Diabetic rats were treated with angrographolide for 2 weeks, then the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in serum were examined and the expressions of Bcl-2 and Box gene of pancreatic cells were detected. RESULTS: Compared with model rats, the plasma glucose levels and the serum MDA contents of the angrographolide-treated rats decreased, serum insulin and activity of SOD increased significatantly (P < 0.01). The expression of Bcl-2 was lower in the model group, while the expression was stronger in the treatment group (P < 0.05). CONCLUSION: Angrographolide can inhibit the apoptosis of islet cells and depress plasma glucose level of diabetic rat model. Its mechanism may be associated with the upregulation of the expression of Bcl-2, the increase of the activity of SOD, the decrease of the production of free radicals and lipid eroxidadion.

Comparison of Epstein-Barr virus genotypes and clinicohistopathological features of nasopharyngeal carcinoma between Guilin, China and Fukuoka, Japan.

Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) is endemic to Guilin, China, but not Fukuoka, Japan. To examine whether the NPC in these two cities are distinct in their association with EBV, we analyzed the histology and EBV genotypes in 163 NPC from Guilin, 52 NPC from Fukuoka and non-cancerous control nasopharyngeal tissues (n=22 each) by in situ hybridization and PCR. The proportion of EBV-positive NPC from Guilin (95%) was higher than that of Fukuoka (55%, p<0.001), and higher in NPC versus control tissues in Guilin (95 vs. 46%) and Fukuoka (55 vs. 32%). Histopathologically, non-keratinizing carcinoma and the undifferentiated subtype (NKC-U) were predominant in Guilin (84%), while the proportion of keratinizing squamous cell carcinoma (KSCC, 38%) was similar to NKC-U (42%) in Fukuoka. EBV-positivity was higher in NKC-U than KSCC in the two cities. EBV genotype A was highly prevalent in Guilin and Fukuoka. However, the BamHI f variant was predominant in Guilin (79%), whereas all the identified types were the F prototype in Fukuoka. The genetic structure and biological functions of the EBV strain associated with endemic NPC in Guilin were probably different from those of Fukuoka. The genetic differences between Guilin and Fukuoka may mirror the differences in histology and patient profiles.