Objective:To study the mRNA expressions of bone morphogenetic protein 2 ( BMP2) in different types of gliomas and the relation between BMP2 mRNA expression and survival time, and to explore the role of BMP2 mRNA expression in prognosis evaluation in gliomas. Methods:The clinical data of 692 patients with gliomas in China Glioma Genome Atlas (CGGA) database were collected. Differences of BMP2 mRNA expression were compared among glioma patients with different histophiologic types, and patients with different gender and different ages, patients at primary or recurrent status, and those with different WHO grading, isocitrate dehydrogenase 1 (IDH1) mutation, 1P19q heterozygous deletion status, and molecular typing. The difference in survival time between patients with high and low BMP2 mRNA expression levels were compared in different categories of gliomas. Results:(1) The BMP2 mRNA expressions were different in different histopathological types of gliomas ( F=9.392, P=0.000); BMP2 expression in the oligodendroglioma subtype was the highest, followed by astrocytoma subtype and glioblastoma. The BMP2 relative mRNA expressions in male and female patients were 9.78±0.65 and 11.26±0.86, respectively, without statistical difference ( P>0.05). The BMP2 relative mRNA expressions in patients <43 years old and patients≥43 years old were 12.51±0.81 and 8.37±0.65, respectively, with significant difference ( P<0.05). The BMP2 relative mRNA expressions were 10.09±0.62 and 10.90±0.93, respectively, without significant difference ( P>0.05). The BMP2 relative mRNA expressions were 13.98±1.12, 12.88±0.88 and 5.18±0.64 in WHO grading II, III, and IV gliomas patients, respectively, with significant differences ( F=30.912, P=0.000). The BMP2 relative mRNA expressions in patients with IDH1 wild-type and IDH1 mutant were 2.73±0.16 and 17.47±0.85, respectively, with significant difference ( P<0.05). The BMP2 relative mRNA expressions in patients with 1P/19Q non-absence and 1P/19Q absence were 7.02±0.36 and 25.28±1.66, respectively, with significant difference ( P<0.05). In patients with lower graded glioma and glioblastoma, the BMP2 mRNA expressions in these patients with IDH mutation were significantly higher than those in patients with IDH wild-type ( P<0.05). (2) In patients with primary glioma and patients with recurrent glioma, the survival time of these patients with high BMP2 mRNA expression (≥4.68) was significantly longer than that of patients with low expression (<4.68, χ2=62.975, P=0.000; χ2=12.810, P=0.000). Conclusion:The BMP2 mRNA expression can be used as an index to predict the malignant degrees of gliomas; patients with high expression have longer survival time than those with low expression.

Objective:To analyze the expression of ATP binding cassette subfamily C member 8 (ABCC8) in different types of gliomas and its relation with overall survival of glioma patients.Methods:The ABCC8 mRNA data and clinical data (gender, age, histopathology, WHO grading, isocitrate dehydrogenase [ IDH] mutation status, 1p/19q deletion status and molecular types), and overall survival of 516 glioma patients from the Cancer Genome Atlas were collected, and the differences of ABCC8 mRNA expression in different types of glioma patients were compared. The differences of overall survival were compared between high (≥54.50) and low (<54.50) ABCC8 mRNA expression patients in different types of glioma patients. Results:(1) ABCC8 mRNA expression in primary glioma patients was significantly higher than that in recurrent glioma patients ( P<0.05). ABCC8 mRNA expression was the highest in oligodendroglioma patients, followed by astrocytoma patients; and glioblastoma patients had the lowest ABCC8 mRNA expression; the differences among glioma patients from subgroups of different histopathological types were statistically significant ( P<0.05). ABCC8 mRNA expression was the highest in patients with grade II glioma, followed by those with grade III ( P<0.05); and patients with grade IV had the lowest ABCC8 mRNA expression ( P<0.05); the differences among patients from subgroups of different WHO grading were statistically significant ( P<0.05). ABCC8 mRNA expression in glioma patients with IDH mutant was significantly higher than that in glioma patients with IDH wild-type ( P<0.05); and ABCC8 mRNA expression in patients with 1P/19Q deletion glioma was significantly higher than that in patients with 1P/19Q deletion ( P<0.05). ABCC8 mRNA expression in low-grade glioma patients and glioblastoma multiforme patients with IDH mutation was significantly higher than that in patients with IDH wild-type ( P<0.05). (2) In all patients with glioma, primary and recurrent glioma, oligodendroglioma, neuroastrocytoma, WHO low-grading (grade II) and WHO high-grading (grade III or IV) glioma, IDH mutation and IDH wild-type gliomas, and 1p/19q deletion and no deletion gliomas, patients with high ABCC8 mRNA expression had significantly higher overall survival time that those with low ABCC8 mRNA expression ( P<0.05). (3) Multivariate Cox analysis showed that ABCC8 mRNA expression was an independent factor for overall survival of patients with gliomas ( HR=0.747, P=0.025, 95%CI:0.579-0.963). Conclusion:The ABCC8 mRNA expression in glioma patients is related to the malignant degrees of gliomas, which can be used as an indicator to predict the prognoses of gliomas.

BACKGROUND: Fibroblast activation protein (FAP) is one of the surface markers of cancer-associated fibroblasts (CAFs) and is closely related to the malignant characterization of CAFs. SP13786 is a specific micromolecule inhibitor of FAP and this study is to investigate the effects and mechanism of SP13786 on the migration and invasion of A549 cells through regulating exosomes of CAFs. METHODS: CAFs and paracancerous fibroblasts (PTFs) were isolated and subcultured from freshly resected lung adenocarcinoma tissues and paracancerous normal tissues separately. MTT assay was used to detect the proliferation of CAFs incubated by different concentrations of SP13786; PTFs-exo, CAFs-exo and CAFs+SP13786-exo were extracted by polymer precipitation method. The A549 cells were divided into Ctrl group, PTFs group, CAFs group and SP13786 group and each group was incubated with DMEM, PTFs-exo, CAFs-exo and CAFs+SP13786-exo separately. Laser confocal microscope was used to observe the endocytoses of exosomes by A549 cells. The expression of alpha-smooth muscle actin (α-SMA) and FAP in PTFs and CAFs and the expression of E-cadherin, N-cadherin, Slug, Stat3 and P-Stat3 in A549 cells were detected by immunofluorescence, immunohistochemistry and Western blot. The migration and invasion ability of A549 cells were detected by cell scratch and transwell methods. RESULTS: α-SMA and FAP were expressed much higher in CAFs than that in PTFs which indicate that CAFs and PTFs were successfully obtained from lung adenocarcinoma and paracancerous tissues (P<0.05). MTT showed that the 50% inhibitory concentration (IC50) of SP13786 for CAFs was about 3.3 nmol/L. In addition, SP13786 can significantly decrease the expression of α-SMA and FAP in CAFs which means that targeted inhibition of FAP could reduce the malignant characteristics of CAFs (P<0.05). Laser confocal microscope found that exosomes from CAFs could be taken up by A549 cells and scratch and transwell tests showed that the endocytosed CAFs-exo could promote the migration and invasion of A549 cells (P<0.001), while FAP inhibitor SP13786 could inhibit the effects of CAFs-exo on A549 cells (P<0.05). Furthermore, Immunofluorescence and Western blot showed that CAFs-exo could promote EMT by decreasing E-cadherin expression and increasing N-cadherin, Slug expression in A549 cells while FAP inhibitor SP13786 could significantly supress CAFs-exo-induced epithelial-mesenchymal transition (EMT) of A549 cells (P<0.05). Moreover, the expression of P-Stat3 was obviously increased in A549 cells of CAFs group and significantly down-regulated in SP13786 group (P<0.05) whereas there was no significant difference in total Stat3 between CAFs and SP13786 groups (P>0.05). Finally, WP1066 (a specific inhibitor of Stat3) was used to comfirm whether SP13786 could influence EMT of A549 cells by inhibiting Stat3 phosphorylation via CAFs-Exo. The results showed that when the phosphorylation of Stat3 in CAFs group was inhibited by WP1066, SP13786 could not influence the P-Stat3 expression and EMT of A549 cells anymore (P>0.05). CONCLUSIONS As a specific micromolecule inhibitor of FAP, SP13786 indirectly inhibits the migration and invasion of A549 cells by affecting exosomes of CAFs. The possible mechanism is to inhibit the phosphorylation of Stat3 and thus affect the EMT of A549 cells.