ACT001 inhibits pituitary tumor growth by inducing autophagic cell death via MEK4/MAPK pathway.

ACT001, derived from traditional herbal medicine, is a novel compound with effective anticancer activity in clinical trials. However, little is known regarding its role in pituitary adenomas. Here, we demonstrated that ACT001 suppressed cell proliferation and induced cell death of pituitary tumor cells in vitro and in vivo. ACT001 was also effective in suppressing the growth of different subtypes of human pituitary adenomas. The cytotoxic mechanism ACT001 employed was mainly related to autophagic cell death (ACD), indicated by autophagosome formation and LC3-II accumulation. In addition, ACT001-mediated inhibitory effect decreased when either ATG7 was downregulated or cells were cotreated with autophagy inhibitor 3-methyladenine (3-MA). RNA-seq analysis showed that mitogen-activated protein kinase (MAPK) pathway was a putative target of ACT001. Specifically, ACT001 treatment promoted the phosphorylation of JNK and P38 by binding to mitogen-activated protein kinase kinase 4 (MEK4). Our study indicated that ACT001-induced ACD of pituitary tumor cells via activating JNK and P38 phosphorylation by binding with MEK4, and it might be a novel and effective anticancer drug for pituitary adenomas.

MicroRNA-93 mediates cabergoline-resistance by targeting ATG7 in prolactinoma.

To date, the management of dopamine agonist (DA)-resistant prolactinomas remains a major clinical problem. Previously, we determined that miRNA-93 expression increases in DA-resistant prolactinomas; however, the role of miRNA-93 in the DA resistance remains largely unexplored. Hence, this study aimed to investigate the susceptibility of tumor cells to cabergoline (CAB) and the autophagy changes in MMQ and GH3 cells after miRNA-93 overexpression or inhibition. We used bioinformatics to identify the potential target of miRNA-93. Subsequently, we analyzed the correlation between miRNA-93 and autophagy-related 7 (ATG7) using protein expression analysis and luciferase assays. Furthermore, the change in the effect of miRNA-93 was measured after ATG7 overexpression. miRNA-93 expression was elevated in DA-resistant prolactinomas, whereas the expression of its identified target, ATG7, was downregulated. miRNA-93 overexpression suppressed the cytotoxic effect of CAB in MMQ and GH3 cells. In contrast, miRNA-93 downregulation enhanced CAB efficiency and promoted cell autophagy, eventually resulting in apoptosis. These results were further confirmed in vivo xenograft models in nude mice. ATG7 overexpression could reverse the inhibitory effect of miRNA-93 on CAB treatment. Taken together, our results suggest that miRNA-93 mediates CAB resistance via autophagy downregulation by targeting ATG7 and serves as a promising therapeutic target for prolactinoma.

Effect of PDGF-Rb antagonist imatinib on endometrial injury repairing in mouse model.

OBJECTIVE: To study the effects of PDGF-Rb antagonists imatinib on endometrial injury repairing in the mouse model. METHODS: The cultured MSCs cells from male mice were marked with BrdU in vitro, and then transplanted to the female mice which suffered from radiation injury through tail vein, PDGF-Rb antagonists imatinib was injected through abdominal cavity. Four groups were arranged, which were radiation transplantation group, normal control group, imatinib intervention group and radiation control group. BrdU incorporation, SRY expression and MVD status were detected in uterus of mice. RESULTS: SRY gene was negative expressed in normal control group and radiation control group. SRY gene presented positive in radiation transplantation group and imatinib intervention group; BrdU incorporation showed negative in radiation control group and normal control group which died in the early stage in mice; the incorporation of BrdU was higher in radiation transplantation group compared with imatinib intervention group; CD34 was positive on the uterus of all the four groups, which showed highest in radiation control group and lowest in radiation control group; The MVD in imatinib intervention group was lower than radiation control group; the difference of MVD was significantly compared with normal control group (P < 0.05). CONCLUSIONS: PDGF-Rb antagonists imatinib could inhibit the repairing function of MSCs in the endometrial lesions in mice.

Relevance of EGFR gene mutation with pathological features and prognosis in patients with non-small-cell lung carcinoma.

OBJECTIVE: To study the relevance of EGFR gene mutation with pathological features and prognosis in patients with non-small-cell lung carcinoma. METHODS: A total of 297 patients from July 2009 to May 2013 were chosen as objects. EGFR gene mutation were detected with fluorescence quantitative PCR. Relevance of EGFR gene mutation with clinical and pathological features was analyzed, and the prognosis of EGFR- mutant-patients and that of EGFR- wide type-patients was compared. RESULTS: In 297 patients, 136 (45.79%) showed EGFR gene mutation. EGFR gene mutation had no significant relevance with age, gender, smoking history, family history of cancer and clinical stage (P>0.05); there was significant relevance between EGFR gene mutation and blood type, pathologic types, differentiation and diameter of cancer (P<0.05). The difference between prognosis of EGFR- mutant-patients and that of EGFR- wide type-patients was statistical significance (P<0.05). CONCLUSIONS: EGFR gene mutation has significant relevance with pathological features, the prognosis of EGFR-mutant-patients is better than that of EGFR- wide type-patients.

Antiproliferative and apoptosis-inducing activity of schisandrin B against human glioma cells.

BACKGROUND: Malignant glioma is the most devastating and aggressive tumour in the brain and is characterised by high morbidity, high mortality and extremely poor prognosis. The main purpose of the present study was to investigate the effects of schisandrin B (Sch B) on glioma cells both in vitro and in vivo and to explore the possible anticancer mechanism underlying Sch B-induced apoptosis and cell cycle arrest. METHODS: The anti-proliferative ability of Sch B on glioma cells were assessed by MTT and clony formation assays. Flow cytometric analysis was used to detect cell cycle changes. Apoptosis was determined by Hoechst 33342 staining and annexin V/PI double-staining assays. The mitochondrial membrane potential was detected by Rhodamine 123 staining. The in vivo efficacy of Sch B was measured using a U87 xenograft model in nude mice. The expressions of the apoptosis-related and cell cycle-related proteins were analysed by western blot. Student's t-test was used to compare differences between treated groups and their controls. RESULTS: We found that Sch B inhibited growth in a dose- and time-dependent manner as assessed by MTT assay. In U87 and U251 cells, the number of clones was strongly suppressed by Sch B. Flow cytometric analysis revealed that Sch B induced cell cycle arrest in glioma cells at the G0/G1 phase. In addition, Sch B induced glioma cell apoptosis and reduced mitochondrial membrane potential (ΔΨm) in a dose-dependent manner. Mechanically, western blot analysis indicated that Sch B induced apoptosis by caspase-3, caspase-9, PARP, and Bcl-2 activation. Moreover, Sch B significantly inhibited tumour growth in vivo following the subcutaneous inoculation of U87 cells in athymic nude mice. COCLUSIONS: In summary, Sch B can reduce cell proliferation and induce apoptosis in glioma cells and has potential as a novel anti-tumour therapy to treat gliomas.

MicroRNA expression profile of bromocriptine-resistant prolactinomas.

MicroRNAs (miRNA) have been implicated in the resistance of tumors to chemotherapy. However, little is known about miRNA expression in bromocriptine-resistant prolactinomas. In this study, 23 prolactinoma samples were classified as bromocriptine-sensitive or -resistant according to the clinical definition of bromocriptine resistance, and their miRNA expression profiles were determined using Solexa sequencing. We found 41 miRNAs that were differentially expressed between the two groups, and 12 of these were validated by stem-loop qRT-PCR. Hsa-mir-93, hsa-mir-17, hsa-mir-22*, hsa-mir-126*, hsa-mir-142-3p, hsa-mir-144*, hsa-mir-486-5p, hsa-mir-451, and hsa-mir-92a were up-regulated and hsa-mir-30a, hsa-mir-382, and hsa-mir-136 were down-regulated in bromocriptine-resistant prolactinomas in comparison with bromocriptine-sensitive prolactinomas. Furthermore, silencing of mir-93 significantly increased the sensitivity of MMQ cells to dopamine agonist treatment. Mir-93 directly affected p21 expression in MMQ cells by targeting the 3'-UTR. Our study is the first to identify a miRNA expression profile associated with bromocriptine-resistant prolactinoma.

Natural history of postoperative nonfunctioning pituitary adenomas: a systematic review and meta-analysis.

OBJECTIVE: Previous studies attempting to define the natural history of postoperative nonfunctioning pituitary adenomas (pNFPAs) were somewhat limited by selection bias and/or small numbers and/or lack of consistency among the study findings. The aim of this study was to scrutinize the literature in order to analyze the natural history of pNFPAs. METHODS: Electronic database including MEDLINE, PubMed and Cochrane CENTRAL were searched. The literature relating to the patients with pNFPAs without postoperative radiotherapy and pharmacotherapy was collected. Eligible studies reported on the rate of tumor recurrence, the tumor growth-free survival rate (TGFSR) at 5 and 10 years, and/or the residual tumor volume doubling time (TVDT). RESULTS: 19 studies met the criteria. The pNFPAs were divided into two groups: the pooled recurrence rate of group I without detectable residual tumor (371 patients) was 12% (95% CI 6-19%), the TGFSR at 5 and 10 years were 96% (95% CI 89-99%) and 82% (95% CI 65-94%), respectively. The pooled recurrence rate of group II with residual tumor (600 patients) was 46% (95% CI 36-56%), the TGFSR at 5 and 10 years were 56% (95% CI 41-71%) and 40% (95% CI 27-53%), respectively. The mean TVDT was 3.4 years (95% CI 2.4-4.5 years). CONCLUSIONS: pNFPAs, with or without detectable residual tumor, need stratification of treatment and radiological/endocrinological follow-up strategy. According to the TVDT, residual tumor regrowth is very slow, which permits an extensive and safe follow-up program for most patients.

[Differential expression analysis of prolactinoma-related microRNAs].

OBJECTIVE: To explore the relationship between the prolactinoma-related microRNAs (miRNA) and the development, growth and hormone secretion of prolactinoma. METHODS: The technique of Solexa sequencing was employed to analyze the differential expressions of prolactinoma and normal anterior pituitary gland samples. And the stem-loop real-time polymerase chain reaction (PCR) was utilized for confirmation. RESULTS: According to the differentially expressed profiles of miRNAs, 4 miRNAs were down-regulated (miR-130a, miR-199b-3p, miR-200b, miR-125b, P < 0.05) and 6 miRNAs up-regulated (miR-342-3p, miR-432, miR-23b, miR-493, miR-493(*), miR-664(*), P < 0.05). The expression levels of miR-493(*) and miR-432 had a significant positive correlation with the serum level of prolactin (r = 0.47, P < 0.05; r = 0.528, P < 0.01) while miR-342-3p a significantly positive correlation with the invasiveness (r = 0.402, P < 0.05). CONCLUSION: miRNAs are differentially expressed between normal anterior pituitary gland and prolactinomas, between invasive and localized prolactinomas and among different hormone secretion levels. It suggests that miRNAs may be involved in the physiological process of development, growth and hormone secretion of prolactinoma.

[Natural history study of postoperative residual non-functioning pituitary adenomas].

OBJECTIVE: To observe the postoperative residual non-functioning pituitary adenomas (PR-NFPAs) without postoperative radiotherapy and to analyze the natural history of PR-NFPAs' growth in order to provide a basis for selecting appropriate strategies of clinical treatment. METHODS: We evaluated the natural history of 20 patients with PR-NFPAs who did not receive postoperative radiotherapy and drug therapy. Through MRI images, the residual tumor volumes of those patients were serially measured. We further calculated the monthly growth rate and the tumor volume doubling time (TVDT) and analyzed the correlations between the patient age, gender, volume of residual tumor, cavernous sinus (CS) invasion and TVDT. RESULTS: All patients received observation alone. Among which, 17 adenomas increased in volume and 3 remained unchanged during a follow-up period of 7 months to 17 years (mean 3.90 yr). The mean patient age was 41.8 years. As to 17 patients with tumor regrowth, the tumor volume at the beginning of MRI observation period was 4.73 cm(3) and tumor volume at the last MRI observation was 16.98 cm(3). During the mean 4-year follow-up period, the average monthly growth rate of PR-NFPAs was 7.87% and the mean TVDT was 724 days. Such factors as patient age, gender, volume of residual tumor and CS invasion did not affect the TVDT of PR-NFPAs. CONCLUSION: The tumor growth rate of PR-NFPAs is not significantly correlated with the patient gender, age, volume of residual tumor and CS invasion. In conjunctions with the volume of PR-NFPAs and the distance between residual adenoma and optic chiasm, we should take the TVDT into consideration and determine the appropriate and safe follow-up period.

Expression of D2RmRNA isoforms and ERmRNA isoforms in prolactinomas: correlation with the response to bromocriptine and with tumor biological behavior.

Invasive prolactinomas are more likely to be resistant to drug therapy but the mechanism of this is still unknown. The objective of this study was to analyze the different expression of ERmRNA and D2RmRNA isoforms in prolactinomas responsive and resistant to dopamine agonist (DA), and to discuss the correlation of such gene expression with tumor biological behavior. A prospective study of 20 consecutive patients who harbored prolactinomas was designed. Patients were classified as responsive (14 cases) or resistant (six cases) according to their clinical and biochemical response to bromocriptine. Tumor tissue samples were examined by means of QRT-PCR analysis. Median D2SmRNA expression in responsive patients was about 2.5-fold that in resistant ones (13.5 +/- 10.4 and 5.4 +/- 2.4, respectively, P = 0.09). No significant difference was found between D2LmRNA expression levels (P = 0.77). However, there was a significant difference between D2S/D2LmRNA ratios for responsive and resistant tumors (P = 0.012). A significant difference was not found between these two groups in levels of ERalphamRNA and ERbetamRNA expression (P = 0.20 and 0.06, respectively). D2SmRNA expression was significantly different for invasive and noninvasive tumors (6.2 +/- 3.6 vs. 17.0 +/- 11.2, respectively, P = 0.02). The D2S/D2L ratio is related to the responsiveness of prolactinomas to DA medication, in which D2SmRNA plays an important role. Lower expression of D2SmRNA in invasive tumor patients suggests that invasive prolactinomas may be more likely to be resistant to DA medication.

Clinical study of simultaneous lung volume reduction surgery during resection of pulmonary or esophageal neoplasms.

BACKGROUND: If the emphysema lesions are not symmetrical, unilateral lung volume reduction surgery (LVRS) can be carried out on the more severe side. The aim of this research was to evaluate the feasibility and effects of LVRS performed simultaneously with resection of pulmonary and esophageal neoplasms. METHODS: Forty-five patients with pulmonary neoplasm and 37 patients with esophageal neoplasm were randomly assigned to group A or group B. In group A, LVRS was performed simultaneously on the same side as thoracotomy. In group B, only tumor resection was performed. The nonfunctional lung area was determined by preoperative chest computed tomography and lung ventilation/perfusion scan. The lung volume removed was about 20% to 30% of the lobes on one side. Preoperative and postoperative indexes including pulmonary function testing variables, arterial blood gas analysis variables, dyspnea scale, 6-minute walk distance, etc., were compared between the groups. RESULTS: There were no surgical deaths in this study. The postoperative forced vital capacity in 1 second, PaO2, PaCO2, dyspnea scale, and 6-minute walk distance were improved significantly in group A, whereas these indexes did not change or decreased slightly in group B. CONCLUSIONS: For tumor patients who have associated emphysema, simultaneous LVRS not only increases the chance of receiving surgical therapy, but also improves the postoperative quality of life of the patient. LVRS has expanded the surgical indication for tumor patients.

[Apoptosis and differentiation induced with TCLA in rat glioblastoma cells in vitro].

OBJECTIVE: To investigate the effect of conjugated trienoic fatty acids on the growth inhibition, apoptosis in rat glioblastoma cells and to elucidate its mechanism of activity. METHODS: Rat glioblastoma cells were tested in vitro cytotoxicity, colony formation inhibition, Brdu incorporation after treatment with TCLA. Its effect of apoptosis induction was detected through Hoechst 33342 staining, cell cycle analysis. The expressions of ADPRTL1, CYP1A1 and PPAR-gamma genes were detected through RT-PCR. RESULTS: After TCLA treatment, the proliferation of C6 cells were inhibited (40 micrommol/L, 72 h, viability 56.71% +/- 0.98%), this action acted as dose-time-dependent relations; colony formation decreased significantly (40 micrommol/L, 0) and BrdU labeling index of cancer cells decreased (63.1% +/- 1.0% vs 95.6 % +/- 1.4%); apoptotic cells increased; By FCM analysis, the apoptotic indices increased, the cells increased in G0/G1 phase, decreased in S phase, which have signigicant difference; RT-RCR showed that TCLA signigicantly increased the level of ADPRTL1, CYP1A1, and PPAR-gamma mRNA expression. CONCLUSION: The findings in this experimental study suggested that TCLA has potent cytotoxicity and induction apoptosis in human and rat glioblastoma cells, its mechanism of activity might be associated with the inhibition of DNA synthesis, cell cycle arrest, up-regulation the expression of apoptosis related genes ADPRTL, CYP1A1, PPAR-gamma.