microRNA-486-5p Regulates DNA Damage Inhibition and Cisplatin Resistance in Lung Adenocarcinoma by Targeting AURKB.

Lung adenocarcinoma (LUAD) severely affects human health, and cisplatin (DDP) resistance is the main obstacle in LUAD treatment, the mechanism of which is unknown. Bioinformatics methods were utilized to predict expression and related pathways of AURKB in LUAD tissues, as well as the upstream regulated microRNAs. qRT-PCR assayed expression of AURKB and microRNA-486-5p. RIP and dual-luciferase experiments verified the binding and interaction between the two genes. CCK-8 was used to detect cell proliferation ability and IC50 values. Flow cytometry was utilized to assess the cell cycle. Comet assay and western blot tested DNA damage and γ-H2AX protein expression, respectively. In LUAD, AURKB was upregulated, but microRNA-486-5p was downregulated. The targeted relationship between the two was confirmed by RIP and dual-luciferase experiments. Cell experiments showed that AURKB knock-down inhibited cell proliferation, reduced IC50 values, induced cell cycle arrest, and caused DNA damage. The rescue experiment presented that high expression of microRNA-486-5p could weaken the impact of AURKB overexpression on LUAD cell behavior and DDP resistance. microRNA-486-5p regulated DNA damage to inhibit DDP resistance in LUAD by targeting AURKB, implying that microRNA-486-5p/AURKB axis may be a possible therapeutic target for DDP resistance in LUAD patients.

The Role of Cavin3 in the Progression of Lung Cancer and Its Mechanism.

OBJECTIVE: The purpose of this study was to describe the role of Cavin3 in the progression of lung cancer and its underlying mechanism. METHODS: Totally, 200 cases of lung cancer tissues and corresponding paracancer tissues were collected. Cavin3 expression in samples was determined by qRT-PCR, and the correlation with lung cancer stages as well as prognosis was statistically analyzed combined with matched clinical information. To investigate the mechanism of Cavin3 in lung cancer progression, firstly, Cavin3 was detected in lung cancer cell lines A549, PC9, and H520. Then, cells with stable Cavin3 overexpression and Cavin3 knockout were established to determine the effect of Cavin3 overexpression on the mammalian target of rapamycin (mTOR) signaling pathway. Subsequently, cells were harvested for cell proliferation, migration, and invasion assays in vitro, as well as nude mouse transplantation tumor experiment in vivo. RESULTS: Cavin3 was seen to be highly expressed in cancer tissues. Statistical analysis with matched clinical data showed that Cavin3 as a prognostic indicator of lung cancer had important clinical value. In addition, it could be found that high expression of Cavin3 was able to promote cell proliferation, migration, and invasion and also potentiate tumor formation in vivo. CONCLUSION: Cavin3 was highly expressed in lung cancer, and it was capable to promote cell proliferation, invasion, and migration.

ZFPM2-AS1 facilitates cell growth in esophageal squamous cell carcinoma via up-regulating TRAF4.

Emerging evidence has confirmed that long noncoding RNAs (lncRNAs) are strongly involved in tumor initiation and development. LncRNA ZFPM2 antisense RNA 1 (ZFPM2-AS1) has been identified as a tumor facilitator in some cancers; nevertheless, its functional significance and regulatory mechanism remain greatly unclear in esophageal squamous cell carcinoma (ESCC). Here, we detected ZFPM2-AS1 expression in ESCC cell lines using qRT-PCR. ZFPM2-AS1 knockdown models were established for investigating the biological function of ZFPM2-AS1 in ESCC cells. The association between miR-3612 and ZFPM2-AS1 or TRAF4 was assessed by RNA pull-down and luciferase reporter assays. The present study indicated that ZFPM2-AS1 was significantly up-regulated in ESCC cells. Functional assays manifested that ZFPM2-AS1 knockdown restrained cell proliferation, migration and invasion, and facilitated cell apoptosis in ESCC. Mechanistically, ZFPM2-AS1 promoted ESCC cell growth and up-regulated TRAF4 to trigger NF-κB pathway by sequestering miR-3612. Besides, miR-3612 was confirmed to be a tumor inhibitor in ESCC. Through restoration experiments, we observed that TRAF4 overexpression could recover the suppressive effect of ZFPM2-AS1 on ESCC cell growth. Collectively, all the results suggested that ZFPM2-AS1 was an oncogene in ESCC cell growth by up-regulating TRAF4 and activating NF-κB pathway.

Effect of miR-21 on Apoptosis in Lung Cancer Cell Through Inhibiting the PI3K/ Akt/NF-κB Signaling Pathway in Vitro and in Vivo.

BACKGROUND/AIMS: Lung cancer is one of the most common malignancies in the world. Apoptosis-stimulating protein of p53 (ASPP2), a tumorigenesis related protein, plays a critical role in the initiation and development of various types of cancers. However, the effect of ASPP2 on lung cancer remains unknown. The purpose of this study aims to investigate the mechanism of ASPP2 regulated by miR-21 in lung cancer in vitro and in vivo. METHODS: In the study, migration and invasion assays, apoptosis assay, caspase activity assay, TUNEL staining, real time PCR and western blot were used to investigate the mechanism of ASPP2 regulated by miR-21 in lung cancer in vitro and in vivo. RESULTS: We demonstrated that the miR-21 inhibitor induced apoptosis through inhibiting the PI3K/Akt/NF-κB signaling pathway in non-small cell lung carcinoma (NSCLC). Moreover, ASPP2 was directly targeted by miR-21 in NSCLC cells. Down-regulation of miR-21 suppressed cell migration and invasion, as well as the EMT signaling pathway in NSCLC cells. Furthermore, the miR-21 inhibitor induced cell apoptosis via the caspase dependent pathway in NSCLC cells. The miR-21 inhibitor enhanced caspase-3, 8, 9 activity in NSCLC cells. In addition, the caspase inhibitor significantly reduced the apoptosis induced by the miR-21 inhibitor in NSCLC cells. CONCLUSIONS: Our results revealed that the miR-21 inhibitor could induce apoptosis through inhibiting the PI3K/Akt/NF-κB signaling pathway in human NSCLC cells, and might serve as a therapeutic strategy to treat NSCLC.

The effects of low-molecular-weight heparin on lung and pulmonary artery injuries in acute pulmonary embolism rat model via platelet-derived growth factor-ß.

OBJECTIVE: To evaluate the effects of anticoagulant agent (low-molecular-weight heparin, LMWH) on the pulmonary artery intima hyperplasia of rats with acute pulmonary embolism (APE) by assaying platelet-derived growth factor-ß (PDGF-ß). METHODS: A total of 90 Sprague-Dawley rats were randomly assigned into the sham, APE, and LMWH groups with 30 rats in each group. The APE rat models were established by injecting autologous blood clots via external jugular veins. In each group, six mice were sacrificed at the 1st day (D1), 4th day (D4), 7th day (D7), 14th day (D14), and 28th (D28) subsequent to the induction of APE to collect the lungs. Right ventricle pressure (RVP) and mean pulmonary arterial pressure (mPAP) were measured. Western blot and RT-PCR analyses were used to assess PDGF-ß expression at various time points. In addition, changes in lung pathology were evaluated using hematoxylin and eosin (H&E) staining and electron microscope. RESULTS: The overall success rate of establishing APE rat models was 85.7% (60/70). There was no difference in mPAP between the sham group and the APE group at the D1, D4, D7, and D14. However, at the D28, mPAP in the APE group was significantly higher than that in the sham group. There was no difference among the three groups regarding RVP. PDGF-ß expression were decreased in the LMWH group at all time points compared with the sham and APE groups (P < 0.01). Furthermore, pulmonary embolism, alveolar wall necrosis and hemorrhage, and inflammation were significantly attenuated in the LMWH group compared with the sham and APE groups subsequent to the induction of APE. CONCLUSION: LMWH attenuates lung and pulmonary artery injuries and improves prognosis. Decreased PDGF-ß in the lungs may be the important factor in the effects observed.

[Protective role of pericardial tissue on injured myocardium in rats].

OBJECTIVE: To define the protective role of pericardial tissue and identify the subcellular population with potential contribution to cardiac protection after injury in a rat model. METHODS: Myocardial infarction (MI) was created by the ligation of left anterior descending artery (LAD) in rats while the pericardial sac was either completely removed (-PC, n = 10) or surgically closed after ligation (+PC, n = 8). The follow-up functional and structure benefits were analyzed by echocardiography and 2,3,5- triphenyl-2H-tetrazolium, chloride(TTC) and bromodeoxyuridine (BrdU) staining. The contributions of cardiac stem cells (sca-1, c-kit and KDR) were evaluated by immunohistochemistry. RESULTS: The integrity of pericardial sac showed significant benefits of cardiac contractile function (ejection fraction and short axis fractional shortening) in comparison to the controls (52 ± 12 vs 37 ± 12 and 36 ± 14 vs 25 ± 13, both P < 0.05). The observed functional amelioration after MI was obvious based on structural repair examined by TTC staining and BrdU incorporation, showing significantly more de novo cardiomyocytes in +PC rats. Interestingly, we have immunohistologically identified a sub-population of KDR+(flk-1+) cells in pericardial tissue which clone genetically distributed and mildly expressed cardiac specific proteins (cTnT). It suggested that the KDR+ cells might act as cardiac progenitors and therefore play an important role in cardiac repair. Meanwhile, rare SCA-1 and c-kit positive cells were detected in pericardial tissue. CONCLUSION: The intact pericardial tissue exerts protective effects after MI. It may be related with a specific cell population of KDR+ cells from pericardial tissue.

[The choice of thoracoscopic sympathectomy in the treatment of palmar hyperhidrosis among different procedures].

OBJECTIVE: Retrospective analyze the long-term efficacy and compensatory sweating of thoracoscopic sympathectomy in the treatment of palmar hyperhidrosis by different operative methods in order to search after a better operative method. METHODS: Retrospective study of 643 cases (498 cases available) palmar hyperhidrosis who accepted video-assisted thoracoscopic bilateral sympathectomy during from 1995 to Aug 2008. The patients were divided into four groups by different operative methods. (1) Group A(n = 82): Thoracoscopic T2-4 sympathectomy was performed. (2) Group B (n = 135): Thoracoscopic T2 sympathectomy was performed. (3) Group C (n = 41): Thoracoscopic T2 sympathetic nerve clipped. (4) Group D (n = 240): Thoracoscopic T3-4 level sympathectomy plus bypass fiber (Kuntz fiber) resection on same level was performed. RESULTS: All procedures were successfully performed under thoracoscope without severe morbidity and mortality. The curative rate of palmar hyperhidrosis was 100.00%. The incidence of compensatory sweating were 54.9% (group A), 48.1% (group B), 48.8% (group C) and 28.8% (group D) respectively with significantly decrease in group D contrast to other three groups. The incidence of high-grade compensatory sweating which have important influences on daily life were 9.8% (group A), 10.4% (group B), 9.8% (group C) and 2.9% (group D) respectively with significantly decrease in group D. Other pairings have nonsignificance. The relapse rate were 1.2% (group A), 2.2% (group B), 7.3% (group C) and 0.8% (group D). Only when group D contrasted to group C has significantly decrease in the relapse rate (χ(2) = 8.423, P = 0.004). Other pairings have nonsignificance. CONCLUSION: The procedure of T3-4 sympathectomy plus bypass fiber resection is reasonable operative method to cure hyperhidrosis with the better curative effect and lowest incidence of compensatory hyperhidrosis.