Inhibition of lncRNA KCNQ1OT1 Improves Apoptosis and Chemotherapy Drug Response in Small Cell Lung Cancer by TGF-β1 Mediated Epithelial-to-Mesenchymal Transition / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#Drug resistance is one of the main causes of chemotherapy failure in patients with small cell lung cancer (SCLC), and extensive biological studies into chemotherapy drug resistance are required. @*Materials and Methods@#In this study, we performed lncRNA microarray, in vitro functional assays, in vivo models and cDNA microarray to evaluate the impact of lncRNA in SCLC chemoresistance. @*Results@#The results showed that KCNQ1OT1 expression was upregulated in SCLC tissues and was a poor prognostic factor for patients with SCLC. Knockdown of KCNQ1OT1 inhibited cell proliferation, migration, chemoresistance and promoted apoptosis of SCLC cells. Mechanistic investigation showed that KCNQ1OT1 can activate transforming growth factor-β1 mediated epithelial-to-mesenchymal transition in SCLC cells. @*Conclusion@#Taken together, our study revealed the role of KCNQ1OT1 in the progression and chemoresistance of SCLC, and suggested KCNQ1OT1 as a potential diagnostic and prognostic biomarker in SCLC clinical management.

Establishment of A Patient-derived Xenotransplantation Animal Model for Small Cell Lung Cancer and Drug Resistance Model / 中国肺癌杂志

BACKGROUND@#Small cell lung cancer (SCLC) is characterized by poor differentiation, high malignancy and rapid growth fast, short double time, early and extensive metastatic malignancy. In clinical, chemotherapy is the main treatment method, while resistance to multiple chemotherapy drugs in six to nine months has been a major clinical challenge in SCLC treatment. Therefore, It has important clinical value to building SCLC aninimal model which is similar to patients with SCLC. Animal model of xenotransplantation (PDX) from the patients with small cell lung cancer can well retain the characteristics of primary tumor and is an ideal preclinical animal model. The study is aimed to establish SCLC PDX animal model and induce the chemoresistance model to help to study the mechanism of chemoresistance and individual treatment.@*METHODS@#Fresh surgical excision or puncture specimens from SCLC patients were transplanted into B-NSGTM mice subcutaneous tissues with severe immunodeficiency in one hour after operation the B-NSGTM mice subcutaneous in 1 hour, and inject chemotherapy drugs intraperitoneally after its tumor growed to 400 mm³ with EP which is cisplatin 8 mg/kg eight days and etoposide 5 mg/kg every two days until 8 cycles. Measure the tumor volum and mice weights regularly, then re-engrafted the largest tumor and continue chemotherapy.@*RESULTS@#Nine cases were conducted for B-NSG mice modeling. Three of nine cases could be engrafted to new B-NSG mice at least two generation. The SCLC PDX animal models have been established successfully. After adopting chemotherapy drugs, the chemoresistance PDX models have been established. High homogeneity was found between xenograft tumor and patient's tumor in histopathology, immunohistochemical phenotype (Syn, CD56, Ki67).@*CONCLUSIONS@#The SCLC PDX animal model and the chemoresistance PDX animal model have been successfully constructed, the success rate is 33%, which provides a platform for the clinical research, seeking for biological markers and choosing individual treatment methods of SCLC.

Role of TGF-β1 in multi-drug resistance in small cell lung cancer and its clinical significance / 中南大学学报(医学版)

Objective:To investigate the role of transforming growth factor β1 (TGF-β1) in multi-drug resistance in small cell lung cancer and its clinical significance.Methods:The mRNA and protein expressions of TGF-β 1 in H69 and H69AR cells were detected by real-time PCR and Western blot,respectively.After silence of TGF-[β1,the sensitivity of H69AR to drugs was detected by CCK8 assay.The expressions of TGF-β1 in lung cancer and paracarcinoma tissues were examined by QRT-PCR and immunohistochemistry.The relationship of TGF-β 1 expression with clinical pathological features and prognosis of patients was studied.Results:Compared to H69,the mRNA and protein expressions of TGF-β1 in H69AR cells were significantly increased by (5.93±0.47) and (8.49±1.92) folds,respectively (P＜0.01).Transfection ofTGF-β1 siRNA resulted in a decrease of TGF-β1 expression by 70.432％ in H69AR ceils (F=21.20,P＜0.01) and an increase insensitivity to chemotherapeutic agents of H69AR cells (t=4.576,P＜0.05).Compare with the paracarcinoma tissues,the expression of TGF-β1 was significantly increased in small cell lung cancer tissues (t=13.925,P＜0.01),which was closely related with clinical stage,chemosensitivity and overall survival (all P＜0.05),but not related with gender,age (both P＞0.05).Conclusion:TGF-β1 is involved in the regulation of small cell lung cancer multidrug resistance,which may be a potential marker to evaluate the chemosensitivity and dinical prognostic for small cell lung cancer.

Expression of miR-139-5p in small cell lung cancer tissue and its clinical significance / 吉林大学学报(医学版)

Objective:To explore the expression of miR-139-5p in small cell lung cancer (SCLC)tissue and its clinical significance, and to clarify the role of miR-139-5p in the occurrence and development of SCLC. Methods:The biological function of miR-139-5p was examined by cell growth,apoptosis and cell cycle analysis. The expressions of miR-139-5p in 50 cases of cancer tissue and paracarcinoma normal tissue were examined by QRT-PCR.Combined with the clinical data,the role of miR-139-5p in clinic was anzlyzed.Results:The expression level of miR-139-5p in SCLC tumor tissue was lower than that in normal lung tissue (P 0.05).The expression level of miR-139-5p in the patient at LD stage was lower than that of the patients at ED stage (P <0.01).The expression level of miR-139-5p in the resistant patients was higher than that of the patients sensitive to chemotherappy (P <0.01).The expression level of miR-139-5p in the survival patients was lower than that in the death patients (P <0.01).Cox regression analysis indicated that miR-139-5p expression and disease stage were the independent prognostic factors for SCLC.Conclusion:miR-139-5p in participates in the occurrence and development of SCLC by inhibiting the cell proliferation,promoting apoptosis and inducing the cell cycle arrest;it may be used as a target gene to evaluate the prognosis of SCLC patients.

Role of SALL4 in regulating multi-drug resistance of small cell lung cancer and its clinical significance / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of SALL4 in regulating multi-drug resistance in small cell lung cancer (SCLC), and to evaluate its clinical significance.</p><p><b>METHODS</b>The expression of SALL4 protein and gene was detected by Western blot and real-time PCR (RT-PCR) in both H69 and H69AR cell lines, respectively. SALL4 expression in H69AR was blocked by the siRNA, and then the drug-sensitivities of H69AR cell lines to chemotherapeutic drugs such as cisplatin, doxorubicin, and etoposide were evaluated by cell counting kit assay. SALL4 expression was also examined by immunohistochemistry, and correlated with patients' clinicopathological features and prognosis.</p><p><b>RESULTS</b>The expression of SALL4 was significantly increased in H69AR cells than in the H69 cells (P < 0.01). Down-regulation of SALL4 increased the drug-sensitivities of H69AR cells to chemotherapeutic drugs (P = 0.02). The expression of SALL4 was significantly increased in SCLC than in para-carcinoma tissues (P < 0.01). SALL4 expression correlated with clinical stage, chemosensitivity and overall survival (P < 0.05), but not with patients' age and gender.</p><p><b>CONCLUSION</b>SALL4 is involved in the regulation of multidrug resistance in SCLC; SALL4 may be a potential target gene to evaluate the chemosensitivity and clinical prognosis for SCLC.</p>

Role of homeobox gene A5 in multidrug resistance of human small cell lung cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of homeobox gene A5 (HOXA5) in multidrug resistance of human small cell lung cancer (SCLC) cells and the possibility of using HOXA5 as the therapeutic targets for SCLC treatment.</p><p><b>METHODS</b>We examined HOXA5 mRNA and protein expressions in chemosensitive human SCLC cells (H69) and the multidrug-resistant SCLC cells (H69AR) using quantitative real-time PCR and immunoblotting. HOXA5 expression was then enhanced or suppressed by transfection of the cells with HOXA5 expression plasmids or small interference RNA (siRNA), and the chemosensitivity of transfected cells to cisplatin (DDP) and etoposide (VP-16) was evaluated using cell counting kit-8 (CCK8) assay.</p><p><b>RESULTS</b>H69 cells showed a 8.99-fold higher expression of HOXA5 than H69AR cells. HOXA5 knockdown caused obvious reductions in the chemosensitivity of H69 cells to DDP and VP-16 with increased cells in G0/G1 phase; conversely, HOXA5 enhancement resulted in an increased sensitivity of H69AR cells to DDP and VP-16.</p><p><b>CONCLUSION</b>HOXA5 may play an important role in multidrug resistance of SCLC and can be a potential therapeutic target in clinical treatment of SCLC.</p>

A Novel Specific Small Molecule Peptide for Non-small Cell Lung Cancer Cell A549 / 生物化学与生物物理进展

To screen small molecule peptide specific binding to non-small cell lung cancer cell (A549) using the "one-bead one-peptide" combinatorial library. Twenty-nine positive beads binding to A549 cell were totally obtained after primary screening. Consensus peptide sequence of -NGXG-was identified by amino acid sequencing in ten beads. Peptide cNGQGEQc was re-synthesized on beads and further studied for its cell specificity, alanine scanning and site-directed deletion. The results showed that cNGQGEQc is specific for cell attachment to non-small cell lung cancer cells including A549, Calu-1 and H178, but not to other tumor cell lines. Both motif of -NGXG-and the length of six peptides are very important for A549 adhesion. Peptide cNGQGEQc labeled with FITC can specifically bind to A549 cell. In a blocking assay with anti-integrin antibodies(?1～6, ?v/?1～5), cell adhesion of A549 to peptide beads was obviously inhibited by integrin ?3 combining with any ? subunits. Results suggested that small molecule peptide cNGQGEQc can bind specifically to non-small cell lung cancer cell A549 via integrin ?3 on cell surface.

A Novel Specific Small Molecule Peptide for Small Cell Lung Cancer / 生物化学与生物物理进展

Screen small molecule peptide specific binding to small cell lung cancer cell (DMS53) was screened by using the "one-bead one-peptide" combinatorial technology. Thirty two positive beads binding to DMS53 were totally obtained after primary screening. Consensus peptide sequences of cXNGRXXc and cNGRXXXc were identified by amino acid sequencing in ten beads. Three representative peptides were re-synthesized on beads. Secondary screening showed that cell adhesion percentage of cFNGRQQc to DMSS was higher than the other two peptides. cFNGRQQc was further studied for cell specificity, alanine scanning and site-directed deletion. The results showed that cFNGRQQc is specific for promoting cell adhesion to DMS53 but not to other human cell lines. Both motif of -NGR- and the length of six peptide of cFNGRQQc structure are important for DMS53 attachment. In an antibody or peptide blocking assay, cell adhesion of DMS53 to peptide bead was not inhibited by antibodies or peptides including anti-integrin, E-cadherin, NCAM and ICAM. The binding site on DMS53 surface for cFNGRQQc peptide need to be proven in the future.

Cell adhesion molecule CD15 and PECAM-1 in renal tissue during acute allograft rejection and their implication / 中华器官移植杂志

The changes in cell adhesion molecule CD15 and PECAM-1 in grafted renal tissue from 18 cases of acute allograft rejection were studied by immunohistochemistry techniques (SP method ) and computerized imaging analysis system. The results showed that: (1) CD15 positive expression level was higher in acute allograft rejection than that in normal renal in tuberular cells.(2) PECAM-1 expression level was lower in glomerular endothelial cells, but higher in tuberular cells than those in normal renal tissue during acute allograft rejection. The above findings suggested that the detection of CD15 and PECAM-1 in the grafted kidney might have some values in the diagnosis of acute allograft rejection.