Oncoprotein SET dynamically regulates cellular stress response through nucleocytoplasmic transport in breast cancer.

SETß is the predominant isoform of oncoprotein SE translocation (SET) in various breast cancer cell lines. Interactome-transcriptome analysis has shown that SETß is intimately associated with cellular stress response. Among various exogenous stimuli, formaldehyde (FA) causes distinct biological effects in a dose-dependent manner. In response to FA at different concentrations, SET dynamically shuttles between the nucleus and cytoplasm, performing diverse biofunctions to restore homeostasis. At a low concentration, FA acts as an epidermal growth factor (EGF) and activates the HER2 receptor and downstream signaling pathways in HER2+ breast cancer cells, resulting in enhanced cell proliferation. Nucleocytoplasmic transport of SETß is controlled by the PI3K/PKCα/CK2α axis and depletion or blockade of the transport of SETß suppresses EGF-induced activation of AKT and ERK. SETß also inhibits not only stress-induced activation of p38 MAPK signaling pathway, but also assembly of stress granules by hindering formation of the G3BP1-RNA complex. Our findings suggest that SET functions as an important regulator which modulates cellular stress signaling pathways dynamically.

SET/PP2A signaling regulates macrophage positioning in hypoxic tumor regions by amplifying chemotactic responses.

Tumor-associated macrophages (TAMs) are one of the main cellular components in the tumor microenvironment (TME). In many types of solid tumors, TAMs tend to accumulate in hypoxic areas and are intimately related to poor patient prognosis. However, the underlying mechanisms by which TAMs infiltrate hypoxic tumor regions remain unclear. In this study, we report that genetic deletion of SE translocation (SET) in myeloid cells inhibited the entry of TAMs into the hypoxic tumor region and abated their proangiogenic and immunosuppressive functions, ultimately inhibiting tumor growth. Mechanistically, in response to hypoxic tumor supernatant stimulation, SET in macrophages shuttled between the nucleus and cytoplasm via the PKC-CK2α signaling axis. Cytoplasmic retention of SET increased ERK and P38 signaling by inhibiting PP2A, which promoted TAM migration into the hypoxic area and polarization toward the M2 phenotype. Therefore, we conclude that SET modulates tumor immunity by acting as a key regulator of macrophage positioning and function in the tumor.

Efficacy and Safety of Regorafenib Monotherapy among Patients with Previously Treated Metastatic Colorectal Cancer in a Chinese Population: A Real-World Exploratory Study.

BACKGROUND: Present study was condeucted to investigate the efficacy and safety of regorafenib for patients with previously treated metastatic colorectal cancer (mCRC) in a Chinese population and the prognostic implications of adverse reactions. METHODS: This retrospective study a total of 96 consecutive patients with mCRC who had failed standard chemotherapy regimens from June 2017 to December 2020. Patients received regorafenib at an initial dosage of 160 mg or 120 mg. The primary end point was progression-free survival (PFS), and secondary end points objective response rate (ORR), disease-control rate (DCR), overall survival (OS), safety, and associations between prognosis and adverse-reaction status. RESULTS: There were three patients with partial response, 49 with stable disease, and 44 with progressive disease. Consequently, the ORR and DCR of the 96 patients were 3.1% (95% CI 0.6%-8.9%) and 54.2% (95% CI 43.7-64.4%), respectively. Prognosis results showed that median PFS of the 96 patients was 2.5 (95% CI 1.98-3.02) months and median OS 9.8 (95% CI 7.02-12.59) months. Additionally, the most frequent adverse reactions during regorafenib treatment were hand-foot syndrome (HFS; 52.1%), hypertension (38.5%), and fatigue (33.3%). Interestingly, the relevance of prognosis to adverse-reaction status exhibited that median PFS of patients with HFS and patients without HFS was 3.3 months and 2.0 months, respectively (P=0.013). Similarly, median PFS of patients with hypertension and without hypertension was 3.6 months and 2.2 months, respectively (P=0.023). CONCLUSION: Potential clinical benefit of regorafenib monotherapy was observed for patients with mCRC who had failed standard chemotherapy regimens. Hypertension and HFS induced by regorafenib therapy could be used as valuable biomarkers to predict the prognosis of regorafenib.

The RNA helicase Dhx15 mediates Wnt-induced antimicrobial protein expression in Paneth cells.

RNA helicases play roles in various essential biological processes such as RNA splicing and editing. Recent in vitro studies show that RNA helicases are involved in immune responses toward viruses, serving as viral RNA sensors or immune signaling adaptors. However, there is still a lack of in vivo data to support the tissue- or cell-specific function of RNA helicases owing to the lethality of mice with complete knockout of RNA helicases; further, there is a lack of evidence about the antibacterial role of helicases. Here, we investigated the in vivo role of Dhx15 in intestinal antibacterial responses by generating mice that were intestinal epithelial cell (IEC)-specific deficient for Dhx15 (Dhx15 f/f Villin1-cre, Dhx15ΔIEC). These mice are susceptible to infection with enteric bacteria Citrobacter rodentium (C. rod), owing to impaired α-defensin production by Paneth cells. Moreover, mice with Paneth cell-specific depletion of Dhx15 (Dhx15 f/f Defensinα6-cre, Dhx15ΔPaneth) are more susceptible to DSS (dextran sodium sulfate)-induced colitis, which phenocopy Dhx15ΔIEC mice, due to the dysbiosis of the intestinal microbiota. In humans, reduced protein levels of Dhx15 are found in ulcerative colitis (UC) patients. Taken together, our findings identify a key regulator of Wnt-induced α-defensins in Paneth cells and offer insights into its role in the antimicrobial response as well as intestinal inflammation.

[Expression of colony-stimulating factor 1 in lung adenocarcinoma and its prognostic implication].

OBJECTIVE: This study aimed to explore the expression of tumor-derived colony-stimulating factor 1 (CSF1), its prognostic significance and underlying related mechanisms in resected lung adenocarcinoma (ADC). METHODS: Immunohistochemistry and tissue microarray were used to detect the expression of CSF1, epidermal growth factor receptor (EGFR), and CD68 in 266 patients with lung adenocarcinoma treated in our department between 2004 and 2008. RESULTS: In the 266 ADC cases, the positive rates of expression of CSF1, EGFR and CD68 proteins were 56.4%, 42.1% and 81.2%, respectively. The expression level of CSF1 was positively correlated with TNM stage, number of involved nodal stations, tumor recurrence and EGFR expression (P<0.05). Univariate analysis indicated that TNM stage, number of involved lymph nodes, number of involved nodal stations, CSF1 expression, the combination of CSF1/EGFR and co-expression of CSF1/CD68/EGFR were statistically significant for prognosis (P<0.05). The results of multivariate analysis showed that TNM stage, co-expression of CSF1/EGFR and CSF1/CD68/EGFR were significant and independent risk factors for survival (P<0.05). Correlational analysis showed that expression of CSF1 and EGFR in the tumors was positively correlated to the degree of infiltration of interstitial tumor-associated macrophages (TAMs) (respectively; P<0.05). CONCLUSIONS: The expression of CSF1 indicates a poor prognosis in postoperative lung adenocarcinoma. Co-expression of CSF1 and EGFR may be a valuable independent prognostic predictor, and its mechanism is probably involved in the interaction of cancer cells and TAMs in the progression of lung adenocarcinoma.

Interstitial tumor-associated macrophages combined with tumor-derived colony-stimulating factor-1 and interleukin-6, a novel prognostic biomarker in non-small cell lung cancer.

OBJECTIVES: Recent experimental evidence has indicated that interstitial tumor-associated macrophages (TAMs), tumor-derived macrophage colony-stimulating factor (also known as CSF-1), and interleukin-6 (IL-6) interact in the pathogenesis of malignant epithelial tumors, including lung cancer. The present study aimed to explore their relationship and prognostic significance in surgically resected non-small cell lung cancer (NSCLC). METHODS: Tissue microarray and immunohistochemistry were used to detect the expression of CSF-1, IL-6, and CD68-positive TAMs in 417 patients with NSCLC undergoing complete pulmonary resection from 2003 to 2008. Their correlations and clinicopathologic data were analyzed using chi-square testing. Their prognostic values were evaluated by univariate Kaplan-Meier survival analysis and multivariate Cox proportional hazard model analysis. RESULTS: The expression of CSF-1 and IL-6 in NSCLC correlated positively with the infiltration degree of TAMs in the tumor stroma (r=0.184 and r=0.196, respectively; P<.001). The expression of both CSF-1 and IL-6 was statistically significant for survival (P<.001). Nevertheless, no such relationship was observed for CD68 in the tumor stroma (P>.05). When CSF-1 and/or IL-6 and CD68 were taken into consideration together, the result became statistically significant. Multivariate analysis showed that co-expression of CD68, CSF-1, and IL-6 remained the most significant and independent prognostic factor for survival (P<.05) but not the combinations of CSF-1 and IL-6, CD68 and CSF-1, or CD68 and IL-6 (P>.05). The 5-year survival rate in the CD68-negative and CSF-1- and IL-6-positive group was better than the rate in the CD68, CSF-1-, and IL-6-positive group (P<.05). CONCLUSIONS: The combination of CD68 plus TAMs, CSF-1, and IL-6 is very likely to be a valuable independent predictor of survival in patients with NSCLC. Perhaps co-expression of CSF-1 and IL-6 induces interstitial TAMs to shift toward the tumor-promoting phenotype.

T1aN0M0 and T1bN0M0 non-small cell lung cancer: a retrospective study of the prognosis.

BACKGROUND: Stage IA of non-small cell lung cancer (NSCLC) is divided into two subgroups, T1aN0M0 (d ≤ 2 cm) and T1bN0M0 (2 < d ≤ 3 cm), in the International Association for the Study of Lung Cancer, seventh edition of TNM Classification of Malignant Tumors. OBJECTIVE: The purpose of this study was the identification of independent clinicopathological predictors of prognosis of these two subgroups of NSCLC. METHODS: Between 1986 and 2005, a cohort of 1,929 cases of stage IA NSCLC in Tian Jin Medical University Cancer Institute and Hospital were retrospectively analyzed. The impact of clinicopathological characteristics on patients' survival was investigated. RESULTS: The overall 5-year survival rate was 71.07%. Patients with T1aN0M0 NSCLC had a better 5-year survival than those with T1bN0M0 (73.98 vs. 68.18%, p = 0.0135). The Cox proportional hazard model revealed that the prognostic factors of T1aN0M0 were intratumoral vessel invasion (p = 0.035) and histologic differentiation (p = 0.004). In patients with T1bN0M0 NSCLC, the prognostic factors were histologic differentiation (p < 0.01), intratumoral vessel invasion (p < 0.01), removal of 6 or more lymph node stations (p < 0.01), and removal of lymph node station 7 (p < 0.01). CONCLUSION: Prognostic factors of T1aN0M0 and T1bN0M0 NSCLC are different. In patients with T1bN0M0 NSCLC, 6 or more lymph node stations and lymph node station 7 should be removed.

Expressions of CLDN1 and insulin-like growth factor 2 are associated with poor prognosis in stage N2 non-small cell lung cancer.

BACKGROUND: Patients with single station mediastinal lymph node (N2) non-small cell lung cancer (NSCLC) have a better prognosis than those with multilevel N2. The molecular factors which are involved in disease progression remain largely unknown. The purpose of this study was to investigate gene expression differences between single station and multilevel N2 NSCLC and to identify the crucial molecular factors which are associated with progress and prognosis of stage N2 NSCLC. METHODS: Gene expression analysis was performed using Agilent 4×44K Whole Human Genome Oligo Microarray on 10 freshfrozen lymph node tissue samples from single station N2 and paired multilevel N2 NSCLC patients. Real-time reverse transcription (RT)-PCR was used to validate the differential expression of 14 genes selected by cDNA microarray of which four were confirmed. Immunohistochemical staining for these validated genes was performed on formalin-fixed, paraffinembedded tissue samples from 130 cases of stage N2 NSCLC arranged in a high-density tissue microarray. RESULTS: We identified a 14 gene expression signature by comparative analysis of gene expression. Expression of these genes strongly differed between single station and multilevel N2 NSCLC. Four genes (ADAM28, MUC4, CLDN1, and IGF2) correlated with the results of microarray and real-time RT-PCR analysis for the gene-expression data in samples from 56 NSCLC patients. Immunohistochemical staining for these genes in samples from 130 cases of stage N2 NSCLC demonstrated the expression of IGF2 and CLDN1 was negatively correlated with overall survival of stage N2 NSCLC. CONCLUSIONS: Our results suggest that the expression of CLDN1 and IGF2 indicate a poor prognosis in stage N2 NSCLC. Further, CLDN1 and IGF2 may provide potential targeting opportunities in future therapies.

Expression of CLDN1 and CLDN10 in lung adenocarcinoma in situ and invasive lepidic predominant adenocarcinoma.

BACKGROUND: Non-mucinous bronchioloalveolar carcinoma (BAC) is considered the early stage of lung adenocarcinoma and is classified as the lung adenocarcioma in situ (AIS) by the International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society. This study was designed to investigate the gene expression differences between AIS (formerly non-mucinous BAC) and invasive lepidic predominant adenocarcinoma (LPA, formerly non-mucinous BAC pattern with >5 mm invasion, mixed type adenocarcinoma with BAC features) and to investigate the mechanism of the progression of lung adenocarcinoma in situ to invasive adenocarcinoma. METHODS: Gene expression analysis was performed by using Agilent 4 × 44 K Whole Human Genome Oligo Microarray on 10 fresh frozen tissue samples of AIS and LPA, respectively. Real time RT-PCR was used to validate the differential expression of 13 genes selected by cDNA microarray on fresh frozen tissue samples from 41 patients with lung adenocarcinoma and 4 genes were confirmed. These 4 genes were then validated by western blotting. Immunohistochemical staining for these validated genes was performed on formalin-fixed, paraffin-embedded tissue samples from 81 cases of lung adenocarcinomna. RESULTS: We identified a 13 gene expression signature by comparative analysis of gene expression. Expression of these genes strongly differed between AIS and LPA. Four genes (MMP-2, c-fos, claudin 1 (CLDN1) and claudin 10(CLDN10)) were correlated with the results of microarray and real time RT-PCR analyses for the gene-expression data in samples from 41 patients with lung adenocarcinoma. As confirmed by western blotting, the expression levels of MMP-2 and c-fos were higher in LPA than those in AIS; the expression levels of CLDN1 and CLDN10 in LPA were lower than those in AIS. Immunohistochemical staining for these genes in samples from 81 cases of lung adenocarcinoma demonstrated the expressions of CLDN1 and CLDN10 were correlated with overall survival of patients with lung adenocarcinoma. CONCLUSIONS: CLDN1 and CLDN10 may play important roles in the development of AIS to LPA. Overexpression of CLDN1 and CLDN10 indicates a favorable prognosis for overall survival in some patients with lung adenocarcinoma. Expression of CLDN10 may be regulated by the c-fos pathway.

Automatic and robust system for correcting microarray images' rotations and isolating spots.

Microarray images contain a large volume of genetic data in the form of thousands of spots that need to be extracted and analyzed using digital image processing. Automatic extraction, gridding, is therefore necessary to save time, to remove user-dependent variations, and, hence, to obtain repeatable results. In this research paper, an algorithm that involves four steps is proposed to efficiently grid microarray images. A set of real and synthetic microarray images of different sizes and degrees of rotations is used to assess the proposed algorithm, and its efficiency is compared with the efficiencies of other methods from the literature.