ARID1A loss induces P4HB to activate fibroblasts to support lung cancer cell growth, invasion, and chemoresistance.

Loss of AT-interacting domain-rich protein 1A (ARID1A) frequently occurs in human malignancies including lung cancer. The biological consequence of ARID1A mutation in lung cancer is not fully understood. This study was designed to determine the effect of ARID1A-depleted lung cancer cells on fibroblast activation. Conditioned media was collected from ARID1A-depleted lung cancer cells and employed to treat lung fibroblasts. The proliferation and migration of lung fibroblasts were investigated. The secretory genes were profiled in lung cancer cells upon ARID1A knockdown. Antibody-based neutralization was utilized to confirm their role in mediating the cross-talk between lung cancer cells and fibroblasts. NOD-SCID-IL2RgammaC-null (NSG) mice received tumor tissues from patients with ARID1A-mutated lung cancer to establish patient-derived xenograft (PDX) models. Notably, ARID1A-depleted lung cancer cells promoted the proliferation and migration of lung fibroblasts. Mechanistically, ARID1A depletion augmented the expression and secretion of prolyl 4-hydroxylase beta (P4HB) in lung cancer cells, which induced the activation of lung fibroblasts through the ß-catenin signaling pathway. P4HB-activated lung fibroblasts promoted the proliferation, invasion, and chemoresistance in lung cancer cells. Neutralizing P4HB hampered the tumor growth and increased cisplatin cytotoxic efficacy in two PDX models. Serum P4HB levels were higher in ARID1A-mutated lung cancer patients than in healthy controls. Moreover, increased serum levels of P4HB were significantly associated with lung cancer metastasis. Together, our work indicates a pivotal role for P4HB in orchestrating the cross-talk between ARID1A-mutated cancer cells and cancer-associated fibroblasts during lung cancer progression. P4HB may represent a promising target for improving lung cancer treatment.

Oncogenic human papillomavirus is not helpful for cytology screening of the precursor lesions of anal cancers in Taiwanese men who are infected with human immunodeficiency virus.

BACKGROUND: Homosexual men infected with human immunodeficiency virus (HIV) are at increased risk of developing anal cancer. The aim of this study was to assess the clinical sensitivity of anal cytology analysis and oncogenic human papillomavirus (HPV) detection for predicting histological anal intraepithelial neoplasia. METHODS: Between March 2011 and December 2013, we enrolled 196 HIV-positive men in Taoyuan General Hospital, Taiwan. We analyzed the results of thin-preparation anal Pap smears, HPV genotyping, and histology of anoscopic biopsy samples. RESULTS: The mean age (±standard deviation) was 31.43 (±8.74) years. The proportion (95 % confidence interval) with abnormal thin-preparation anal cytology was 36.2 % (29.8-43.2 %): 16.8 % (12.2-22.7 %) atypical squamous cells of undetermined significance, 14.8 % (10.5-20.4 %) low-grade squamous intraepithelial lesions, and 4.6 % (2.4-8.5 %) high-grade squamous intraepithelial lesions. At least one HPV genotype was detected in 90.8 % of subjects, and the mean number of HPV infection types was 4.41 (±3.24).The frequency of histological high-grade anal intraepithelial lesions was 7.14 % (95 % confidence interval 4.3-11.6 %). Anal cytology yielding atypical squamous cells of undetermined significance or higher grades resulted in a sensitivity of 64.3 %, specificity of 65.9 %, positive predictive value of 12.7 %, and negative predictive value of 96 %. Using both oncogenic HPV and cytology did not provide better performance. CONCLUSIONS: Anal cytology yielding atypical squamous cells of undetermined significance or higher grades could detect two-thirds of high-grade anal intraepithelial neoplasias in HIV-infected men and should be promoted for anal cancer prevention.

Screening and risk factors for anal cancer precursors in men infected with HIV in Taiwan.

Homosexual men infected with human immunodeficiency virus (HIV) are at a greater risk of developing anal cancer. Men who are infected with HIV and visited the outpatient clinics in Taoyuan General Hospital were enrolled to this study. During March to December 2011, thin preparation anal Pap smear and human papillomavirus (HPV) genotyping were performed in 230 subjects, of which 69 subjects underwent anoscopic biopsy. Their mean age was 32.9 ± 8.1 years, and 181 (78.6%) men were homosexual. The proportion and 95% confidence interval (CI) of subjects with anal dysplasia in cytology was 23.0% (17.56-28.44), including 13.4% (9.26-18.14) with atypical squamous cells of undetermined significance, 7.0% (3.70-10.30) with low-grade squamous intraepithelial lesions, and 2.6% (0.54-4.66) with high-grade squamous intraepithelial lesions. For participants having atypical squamous cells of undetermined significance or higher grades, multivariate logistic regression models yielded adjusted odds ratios (95% CI) of 12.61 (1.63-97.56) for homosexuality, 1.62 (1.31-2.00) for number of oncogenic HPV types, and 1.01 (1.00-1.02) for number of lifetime sexual partners. For detection of histological grade II or III anal intraepithelial neoplasm in anoscopic biopsies, the sensitivity of sequential tests for oncogenic HPV and cytology with atypical squamous cells of undetermined significance or higher grades was 100%. The positive likelihood ratio was 3.09 (P = 0.05). It is important to consider anal cancer precursors among homosexual men who are infected with HIV. Anal cytology and oncogenic HPV genotyping testing are effective screening methods.

PLAU promotes growth and attenuates cisplatin chemosensitivity in ARID1A-depleted non-small cell lung cancer through interaction with TM4SF1.

Loss of ARID1A, a subunit of the SWI/SNF chromatin remodeling complex, contributes to malignant progression in multiple cancers including non-small cell lung cancer (NSCLC). In the search for key genes mediating the aggressive phenotype caused by ARID1A loss, we analyzed 3 Gene Expression Omnibus (GEO) datasets that contain RNA sequencing data from ARID1A-depleted cancer cells. PLAU was identified as a common gene that was induced in different cancer cells upon ARID1A depletion. Overexpression of PLAU positively modulated NSCLC cell growth, colony formation, cisplatin resistance, and survival under serum deprivation. Moreover, enforced expression of PLAU enhanced tumorigenesis of NSCLC cells in nude mice. Mechanistically, PLAU interacted with TM4SF1 to promote the activation of Akt signaling. TM4SF1-overexpressing NSCLC cells resembled those with PLAU overepxression. Knockdown of TM4SF1 inhibited the growth and survival and increased cisplatin sensitivity in NSCLC cells. The interaction between PLAU and TM4SF1 led to the activation of Akt signaling that endowed ARID1A-depleted NSCLC cells with aggressive properties. In addition, treatment with anti-TM4SF1 neutralizing antibody reduced the growth, cisplatin resistance, and tumorigenesis of ARID1A-depleted NSCLC cells. Taken together, PLAU serves as a target gene of ARID1A and promotes NSCLC growth, survival, and cisplatin resistance by stabilizing TM4SF1. Targeting TM4SF1 may be a promising therapeutic strategy for ARID1A-mutated NSCLC.

Gene Targets Network Analysis for the Revealing and Guidance of Molecular Driving Mechanism of Lung Cancer.

The objective was to explore the function of gene differential expressions between lung cancer tissues and the interaction between the relevant encoded proteins, thereby analyzing the important genes closely related to lung cancer. A total of 120 samples from the GEO database (including two groups, i.e., 60 lung cancer in situ specimens and 60 normal specimens) were taken as the research objects, which were submitted to the analysis of signaling pathway, biological function enrichment, and protein interactions to reveal the molecular driving mechanism of lung cancer. Results: A total of 875 differentially expressed genes were obtained, including 291 up-regulated genes and 584 down-regulated genes. The up-regulated genes were mainly involved in biological processes such as protein metabolism, protein hydrolysis, mitosis, and cell division. Down-regulated genes were mainly involved in neutrophil chemotaxis, inflammatory response, immune response, and angiogenesis. The protein expression of high expression genes and low expression genes in patients were higher than those in the control group. The protein corresponding to the high expression gene was highly expressed in the patient group. Meanwhile, the proteins corresponding to the low expression genes were also expressed in the patient group, which showed that although the proteins corresponding to the low expression genes were low in the patients, they were still the target genes related to lung cancer. In conclusion, the molecular driving mechanism in lung cancer was mainly related to protein metabolism, proteolysis, mitosis, and cell division. It was found that TOP2A, CCNB1, CCNA2, CDK1, and TTK might be the critical target genes of lung cancer.

Diffuse expression of RNA-binding protein IMP3 predicts high-stage lymph node metastasis and poor prognosis in colorectal adenocarcinoma.

BACKGROUND: IMP3 (insulinlike growth factor II mRNA-binding protein 3) is a newly identified oncofetal RNA-binding protein that is involved in cell growth and cell migration during the early stages of embryogenesis. This study sought to elucidate its role in tumor progression and prognosis of colorectal adenocarcinoma (CRA). METHODS: IMP3 expression in 186 surgically resected unifocal primary CRAs was analyzed by immunohistochemistry. The proportions of tumor cells positive for IMP3 were scored into diffuse (> or =50%), focal or heterogeneous (10-50%), and trace (<10%), and the expression levels were correlated with clinicopathologic features and patient survival. RESULTS: Cytoplasmic immunoreactivity for IMP3 was diffuse in 66 (35%), focal or heterogeneous in 38 (21%), and trace in 34 (18%) samples. No staining was seen in the adjacent nontumorous tissue. Diffuse IMP3 expression correlated with large tumor (>3 cm, P = .0452), high-stage tumor (IIIa-IV, P = .0417), lymph node metastasis (P = .0232), high lymph node ratio (LNR > or = .7, P = .0016), and lower 5-year survival (P = .0012). Further analysis showed that patients with high-stage CRA and diffuse IMP3 expression had the worst survival rate (P < .0001)-far worse than those without diffuse IMP3 expression (P = .0038). Moreover, multivariant analysis showed diffuse IMP3 expression, serosal invasion, LNR, tumor stage, and adjuvant chemotherapy were independent prognostic factors in CRA. CONCLUSION: Diffuse IMP3 protein expression correlates with invasion and aggressiveness during cancer growth and metastasis, and it is an important prognostic factor of CRAs.

Function of CSE1L/CAS in the secretion of HT-29 human colorectal cells and its expression in human colon.

The secretion of colorectal epithelium is important for maintaining the physiological function of colorectal organ. Herein, we report that cellular apoptosis susceptibility (CAS) (or CSE1L) protein regulates the secretion of HT-29 human colorectal cells. Polarity is essential for directed secretion of substances produced by epithelial cells to the external (luminal) compartment; CAS overexpression induced polarization of HT-29 cells. CAS was punctate stained in the cytoplasm of HT-29 cells, and CAS overexpression increased the translocation of CAS-stained vesicles to the cytoplasm near cell membrane and cell protrusions. CAS overexpression increased the secretion of carcinoembryonic antigen (CEA) and cathepsin D. Immunohistochemistry showed CAS was positively stained in the goblet cells of colon mucosa and cells in the crypts of Lieberkühn of human colon as well as the glands in metastatic colorectal cancer tissue. Our results suggest that CAS regulates the secretion of colorectal cells and may regulate the metastasis of colorectal cancer.

Human papillomavirus infection and anal cytology in Taiwanese homosexual men with and without HIV infection.

INTRODUCTION: Anal cancer screening has not been adopted by Taiwanese care providers. The study aim was to explore the differences of anal cytology and HPV detection among men with and without HIV. METHODOLOGY: In this case-control study, men with HIV who attended one of the outpatient clinics of Taoyuan General Hospital were enrolled as cases. Men who had experienced condomless sex and tested HIV negative were enrolled as controls. Anal swabs were collected for thin-preparation anal cytology and HPV genotyping. RESULTS: A total of 288 men who had tested positive for HIV and 208 who had tested HIV negative were enrolled; 75% of subjects with HIV and 30.3% of those without HIV had tested positive for various types of HPV (P < 0.001). Anal cell dysplasia, including atypical squamous cells with undetermined significance (ASCUS), low-grade squamous intraepithelial lesions (LSILs), high-grade squamous intraepithelial lesions (HSILs), or atypical squamous cells cannot exclude HSIL (ASC-H), were noted in 20.8% of men with HIV and 4.8% of those without HIV (P < 0.001). In multivariate analysis, HIV serostatus, history of sexually transmitted infections, having male sexual partners, and practice of anal sex were correlated significantly with detection of any type of HPV. Additionally, both oncogenic and non-oncogenic HPV types were significantly associated with anal cytology dysplasia. CONCLUSIONS: We strongly suggest that there should be awareness of anal HPV infection and related anal cellular dysplasia in at-risk populations.

Multiple types of human papillomavirus infection and anal precancerous lesions in HIV-infected men in Taiwan: a cross-sectional study.

OBJECTIVES: This study aimed to assess the relationship between infection with multiple human papillomavirus (HPV) types and abnormal anal cytology in HIV-infected men. DESIGN: An observational, cross-sectional study. SETTING: A regional referral hospital in Taiwan. PARTICIPANTS: In total, 714 HIV-infected men were enrolled between March 2011 and June 2016. Thin preparation anal Pap smears were interpreted according to the 2001 Bethesda System. Thirty-seven types of HPV were detected by reverse line blotting, including 13 oncogenic types and 24 non-oncogenic types. OUTCOME MEASURES: The relationship between anal HPV infection and abnormal anal cytology in people of Asian ethnicity and the coverage efficacy in HPV-vaccinated HIV-infected men. RESULTS: On anal cytology, 175 (24.5%) subjects had atypical squamous cells of undetermined significance (ASCUS) or higher grades of dysplasia, including 87 (49.7%) with ASCUS, 73 (41.7%) with low-grade squamous intraepithelial lesions (LSILs) and 15 (8.6%) with high-grade squamous intraepithelial lesions (HSILs). A higher proportion of subjects with those without LSIL/HSIL (93.1% vs 67.3%, P<0.0001) had multiple HPV types. The odds of having LSIL/HSIL increased with an increasing number of HPV types: the ORs ranged from 1 for no HPV types to 6.96 (95% CI 2.38 to 20.37) for more than five types (Ptrend <0.0001). Multivariate logistic regression analysis showed a significant association between LSIL/HSIL and the number of HPV genotypes present (OR 1.20; 95% CI 1.02 to 1.42, P<0.05). HPV types covered by the nonavalent HPV vaccine (types 6/11/16/18/31/33/45/52/58) were detected in 70.1% of the patients in this study. CONCLUSIONS: The odds of having anal LSIL/HSIL are approximately seventimes greater in HIV-infected men with than withoutsix or more types of HPV. Multiple HPV types in HIV-infected patients deserves aggressive follow-up, and HPV vaccination programme require scaling up.

Electroencephalographic features of patients with SCN1A-positive Dravet syndrome.

OBJECTIVE: The aim of this study was to characterize the awake EEG features of patients with SCN1A-positive Dravet syndrome. METHODS: Between January 2002 and December 2012, clinical data of 37 SCN1A-positive Dravet syndrome patients were collected. The first interictal awake EEG features, hot water bath test induced ictal seizure patterns and the concomitant EEG results, as well as follow-up interictal awake EEG recordings were analyzed. RESULTS: Thirty-seven interictal awake EEG recordings showed 43.2% had normal features, 43.2% had nonspecific findings, and 13.5% had abnormal epileptiform discharges. Ictal pleomorphic seizure types with a median number of three were recorded in 26 patients. In total, 42.3% exhibited myoclonic seizures as their first recognizable seizure type with simultaneous EEG findings characterized by generalized or focal spikes, generalized 2-3.5Hz spike and wave discharges, or generalized 2-3Hz high voltage slow waves, and 30.8% manifested atypical absence seizures with concomitant EEG results showing generalized or focal spikes. Fifteen patients had 45 follow-up interictal awake EEGs during a period of six years. The follow-up awake EEG recordings revealed 42.2% had normal features, 42.2% showed nonspecific findings, and 15.6% disclosed epileptiform discharges. CONCLUSIONS: The initial and follow-up interictal awake EEG recordings showed normal results and nonspecific features in the majority of SCN1A-positive Dravet syndrome patients. Ictal electroencephalographic seizure types and concomitant EEG pictures were quite diverse and polymorphous. A low detection rate of interictal epileptiform abnormalities at awake stage might make patient management more challenging.