Shunchang Jiao

Efficacy analysis of tyrosine kinase inhibitors on rare non-small cell lung cancer patients harboring complex EGFR mutations

The efficacy of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKI) in patients with non-small cell lung cancer (NSCLC) is related to EGFR mutations. Although the p.L858R point mutation in exon 21 and the in-frame deletion mutation in exon 19 are well known, efficacy of EGFR-TKI in patients with more than one EGFR mutation is not well understood. 799 NSCLC patients were screened for EGFR mutations. Of the 799 patients, 443 (55.4%) had mutations, out of which 22 (2.75%) had multiple complex mutations. Most multiple mutations (20/22) harbored common mutations such as the p.L858R point mutation in exon 21 and the in-frame deletion mutation in exon 19. 11 out of 22 patients who had multiple EGFR mutations underwent TKI therapy and primary end-points of progression free and overall survival were determined. Our analysis revealed that cases with multiple mutations had similar end-point outcomes as single mutation to TKI therapy. Report of these cases will be helpful in decision making for treatment of NSCLC patients harboring multiple EGFR mutations.

ILEI drives epithelial to mesenchymal transition and metastatic progression in the lung cancer cell line A549.

Transforming growth factor beta (TGF-β) induces epithelial–mesenchymal transition (EMT) accompanied by cellular differentiation and migration. Despite extensive transcriptomic profiling, identification of TGF-β-inducible, EMT-specific genes during metastatic progression of lung cancer remains elusive. Here, we functionally validate a previously described post-transcriptional pathway by which TGF-β modulates expression of interleukin-like EMT inducer (ILEI), and EMT itself. We show that poly r(C)-binding protein 1 (PCBP1) binds ILEI transcript and repress its translation. TGF-β activation leads to phosphorylation at serine-43 of PCBP1 by protein kinase Bβ/Akt2, inducing its release from the ILEI transcript and translational activation. Modulation of hnRNP E1 expression modification altered TGF-β-mediated reversal of translational silencing of ILEI transcripts and EMT. Furthermore, ILEI could induce, as well as maintain, CD24lowCD44high subpopulation in A549 cells treated with TGF-β, which might explain its capability to induce metastatic progression. These results thus validate the existence of an evolutionary conserved TGF-β-inducible post-transcriptional regulon that controls EMT and subsequent metastatic progression of lung cancer.

Efficacy of nimotuzumab plus gemcitabine usage as first-line treatment in patients with advanced pancreatic cancer.

Advanced pancreatic cancer patients have poor prognosis and scarcely respond to conventional therapies. Clinical trials support the use of molecular-targeted therapy against epidermal growth factor receptor (EGFR) signaling. The objective of the current study was to evaluate the contribution of a monoclonal antibody against EGFR, nimotuzumab, to standard gemcitabine therapy. Patients with unresectable locally advanced or metastatic pancreatic adenocarcinoma were assigned to receive gemcitabine plus nimotuzumab. The primary end point was overall survival, whereas the secondary end points included progression-free survival, objective response, and adverse side effects. A total of 18 eligible patients were accrued between December 2007 and July 2010. The disease control rate, calculated as the sum of complete response, partial response, and stable disease, was 55.6xa0%. The median overall survival time was 9.29xa0months (95xa0% CI, 5.499 to 13.072). The median progression-free survival was 3.71xa0months (95xa0% CI, 2.526 to 4.902), and the 1-year survival rate was 38.9xa0%. Of all the patients, 88.8xa0% had at least one adverse side effect; however, no grade 4 adverse side effect was reported. Nimotuzumab as a high-purity humanized monoclonal antibody with favorable safety profile, its value in the treatment of pancreatic cancer along with gemcitabine, particularly in the comprehensive treatment of advanced pancreatic cancer, is appealing for further prospective randomized large-scale clinical trials.

Effects of ARHI on breast cancer cell biological behavior regulated by microRNA-221.

The aplysia ras homolog member I (ARHI) is a tumor suppressor gene and is downregulated in various cancers. The downregulation of ARHI was regulated by miR-221 in prostate cancer cell lines. However, it has not been reported whether ARHI is regulated by miR-221 in breast cancer. Here, we reported that the ARHI protein level was downregulated in breast cancer tissues and breast cancer cell lines. The overexpression of ARHI could inhibit cell proliferation and invasion and induce cell apoptosis. To address whether ARHI is regulated by miR-221 in breast cancer cell lines, the results in this study showed that a significant inverse correlation existed between ARHI and miR-221. MiR-221 displayed an upregulation in breast cancer tissues and breast cancer cell lines. The inhibition of miR-221 induced a significant upregulation of ARHI in MCF-7 cells. To prove a direct interaction between miR-221 and ARHI mRNA, ARHI 3′UTR, which includes the potential target site for miR-221, was cloned downstream of the luciferase reporter gene of the pMIR-REPORT vector to generate the pMIR-ARHI-3′UTR vector. The results confirmed a direct interaction of miR-221 with a target site on the 3′UTR of ARHI. In conclusion, ARHI is a tumor suppressor gene that is downregulated in breast cancer. The overexpression of ARHI could inhibit breast cancer cell proliferation and invasion and induce cell apoptosis. This study demonstrated for the first time that the downregulation of ARHI in breast cancer cells could be regulated by miR-221.

Dysregulation of Circulating Follicular Helper T Cells in Nonsmall Cell Lung Cancer

Nonsmall cell lung cancer (NSCLC) is greatly affected by the dysregulation of the immune system. Circulating follicular helper T cells (Tfh) play critical roles in inducing B-cell activation and producing various cytokines. In the current study, we investigated levels of circulating Tfh in NSCLC. Circulating Tfh and it subtypes were determined by measuring CD3, CD4, CXCR5, CXCR3, and CCR6 in 62 NSCLC patients and 66 healthy controls using flow cytometry. Data presented that percentage of circulating Tfh in the peripheral CD4(+) T cells was significantly increased in NSCLC (14.0%) than in controls (8.7%) (p<0.001). Further analysis revealed that the upregulation of Tfh was contributed by the Th2-Tfh subtype and the Th17-Tfh subtype, whereas the percentage of the Th1-Tfh subtype was significantly decreased in patients. Investigating the clinical stages of the patients demonstrated that prevalence of Tfh was significantly elevated in cases with advanced stages (III: 14.2%; IV: 16.4%) than those with primary stages (I: 10.9%; II: 10.8%). In addition, we analyzed Tfh in patients with different histological types. Results showed that the percentage of circulating Tfh was further upregulated in adenocarcima than squamous cell carcinoma or other types. This study suggests the involvement of circulating Tfh in the pathogenesis and progression of NSCLC, and provides a potential pathway for understanding this disease.

The effect of lysyl oxidase polymorphism on susceptibility and prognosis of nonsmall cell lung cancer

Lysyl oxidase (LOX) is a copper-dependent amine oxidase that plays important roles in development and homeostasis of the lungs. The aim of this study was to investigate whether polymorphisms in the LOX gene were associated with susceptibility to nonsmall cell lung cancer (NSCLC). We sequenced the promoter region of LOX gene and tested the previously reported polymorphism 473xa0G/A in the Han Chinese population. A novel polymorphism, −22xa0G/C, was identified in the promoter region of LOX. However, it did not show any correlation with NSCLC. Frequencies of the 473AA genotype and 473A allele were significantly higher in the NSCLC cases than in control group (pu2009=u20090.004 and pu2009=u20090.006, respectively). Further, our results showed that survival time of NSCLC patients with 473AA genotype was significantly shorter compared to the cases carrying 473xa0G allele (20.0xa0months vs. 28.0xa0months, pu2009=u20090.011). These data indicate that LOX 473xa0G/A polymorphism is associated with increased risk of NSCLC and can be a prognostic predictor for this disease.

Effects of ARHI on cell cycle progression and apoptosis levels of breast cancer cells

The purposes of this study were to investigate the role of Aplysia Ras Homolog I (ARHI) on cell growth, proliferation, apoptosis, and other biological characteristics of HER2-positive breast cancer cells. Our goal was to provide experimental evidence for the development of future effective treatments of HER2-positive breast cancer. A pcDNA3.1-ARHI eukaryotic expression vector was constructed and transfected into the human HER2-positive breast cancer cell lines SK-BR-3 and JIMT-1. Then, various experimental methods were utilized to analyze the biological characteristics of ARHI-expressing breast cancer cells and to examine the impact of expression of the ARHI gene on cyclin D1, p27Kip1, and calpain1 expression. We further analyzed the cells in each group after treatment with trastuzumab to examine the effects of this drug on various cellular characteristics. When we compared pcDNA3.1-ARHI-expressing SK-BR-3 and JIMT-1 cells to their respective empty vector and control groups, we found that cell viability was significantly lower (pu2009<u20090.05) in the ARHI-expressing cells, and the proportions of G1 phase cells and apoptotic cells were significantly higher in the ARHI-expressing cells (pu2009<u20090.05). In all groups of SK-BR-3 cells, trastuzumab treatment significantly decreased cell growth (pu2009<u20090.05). The proportion of cells in G1 phase and the number of apoptotic cells in the pcDNA3.1-ARHI-expressing group were significantly higher than that in the empty vector group and the control group (pu2009<u20090.05). The growth of pcDNA3.1-ARHI-transfected JIMT-1 cells was significantly decreased (pu2009<u20090.05), while the proportion of apoptotic cells was significantly increased (pu2009<u20090.05). Cell growth, viability, and the percentage of apoptotic cells were similar between the JIMT-1 empty vector and control groups. ARHI expression inhibited cyclin D1 expression in SK-BR-3 cells and JIMT-1 cells, while it promoted p27Kip1 and calpain1 expression in these cells. ARHI expression inhibits the growth and proliferation of HER2-positive breast cancer cells, while it also promotes apoptosis in these cells. ARHI expression also improves the sensitivity of JIMT-1 cells to trastuzumab by inducing apoptosis.

Activating transcription factor 3 promotes colon cancer metastasis

Colorectal cancer is one of the commonest of solid malignancy in the world. Activating transcription factor 3 (ATF3), a homolog of the mouse TI-241 and rat LFR-1, is a stress responsive gene that has been widely indicated in different malignancies. However, the role of ATF3 in colon cancer is paradoxical with both a suggested pro- and anti-tumorigenic role. The objective of the current study was to investigate the role of ATF3 in colon cancer metastasis using HT29 and CaCO2 colon cancer cell lines. Expression of ATF3 was initially evaluated in five pairs of colon cancer and matched noncancerous colon tissues. The role of ATF3 in promoting in vitro migration and invasion were evaluated by siRNA-mediated knockdown and adenovirus-mediated overexpression of ATF3. In addition, the role of ATF3 in promoting in vivo tumor growth and hepatic metastasis was investigated by shRNA-mediated knockdown of ATF3. Expression of ATF3 was more in the colon cancer tissues as compared with the pooled noncancerous control colon tissue. Our results showed that in both HT29 and CaCO2 cells, ATF3 promoted in vitro motility and invasion. Furthermore, knockdown of ATF3 attenuated subcutaneous tumor growth and CD31+ neovasculature in xenograft assays with HT29 and CaCO2 cells and inhibited hepatic metastasis. Cumulatively, our results unequivocally show that ATF3 promotes colon cancer metastasis.

GINS2 regulates matrix metallopeptidase 9 expression and cancer stem cell property in human triple negative Breast cancer.

GINS2, a subunit of GINS complex, is critical for the initiation of DNA replication and DNA replication fork progression. The expression of GINS2 is misregulated in many malignant tumors, such as leukemia, breast cancer and melanoma. However, the role of GINS in breast cancer remains poorly characterized. We investigate the possible effect and particular mechanism of GINS in breast cancer cells. We showed that expression of GINS2 is enriched in triple negative breast cancer (TNBC) cell lines. Furthermore, GINS2 knockdown decreased the growth, invasive ability and stem-like property of TNBC cells. Mechanistically, silencing of GINS2 in TNBC cells caused dramatic decrease of matrix metalloproteinase-9 (MMP9). Finally, the abundance of GINS2 correlated with the advance stages of tumor in human TNBC patients. Our studies provided insight into the molecular regulation of TNBC progression and invasion. More importantly, our data suggest that GINS2 could be an outstanding therapeutic target for inhibiting invasive TNBC growth and metastasis.

Detection of circulating tumour cells in gastric and hepatocellular carcinoma: a systematic review.

Objective This systematic review was conducted to summarize the use of circulating tumour cell (CTC) detection as a prognostic indicator in gastric cancer and hepatocellular carcinoma (HCC). Methods Databases (MEDLINE®, EMBASE®, SCOPUS, Web of Science, Conference Proceedings Citation Index-Science, Database of Abstracts of Reviews of Effects, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov) were searched to identify studies that reported the detection of CTCs in patients with gastric cancer or HCC. Results Fifteen studies in patients with gastric cancer and 10 studies in patients with HCC, with a total of 793 and 577 patients, respectively, met the specific inclusion criteria for further analysis. Heterogeneity and potential bias among the studies prevented any statistical analysis. Conclusion Several methodological techniques have allowed the detection of CTCs in patients with gastric cancer or HCC, but the studies identified in this report generally reported on small cohorts and there was heterogeneity and potential bias in the studies. This highlights the need for large systematic multicentre clinical trials to confirm the potential prognostic benefit of detecting CTCs in patients with cancer.