Effect of TWIST1 Gene on the Proliferation and Apoptosis of Human Glioma Cell Line TJ861 by Regulating Mammalian Target of Rapamycin Signaling Pathway

To observe TWIST1 gene expression in human glioma and study the effect human glioma cell line TJ861 on the proliferation and apoptosis, and further explore its potential mechanism to provide some reference for the targeted treatment of glioma in the future. Detection of cancer tissue (Carcinoma tissue) in 55 patients with glioma by RT-PCR and Expression level of TWIST1 in normal and paracancerous tissues (Adjacent tissue), the human glioma cell line TJ861 was further divided into, Nonsense sequence group, (si-NS group), TWIST1 Inhibition group (si-TWIST1 group) and control group. The glioma cells of si-NS group and si-TWIST1 group were transfected with nonsense sequence and TWIST1 siRNA respectively by liposome transfection technology. Use CCK8 assay to test the cell proliferation ability of each group at 0, 12, 24, 36, 48 and 72 hours; 48 hours after siRNA transfection, The ability of DNA replication in each group was detected by EdU staining; Apoptosis related protein expression, in each group, was analyzed by Western blot; TUNEL staining was used to test the apoptosis rate of each group; In the end, We studied TWIST1 effect knocking down on mTOR protein expression in human glioma cells and mTOR protein expression in cancer and adjacent tissues. TWIST1 expression in glioma cells was higher, compared with normal tissues (P <0.05); After transfection of TWIST1 siRNA into human glioma cell line TJ861 in vitro, CCK8 showed glioma cells proliferation ability in si-TWIST1 group at 12, 24, 36, 48 and 72 hours was lower, compared with the control group (P <0.05); After siRNA transfection at 48 hours, the DNA replication ability of glioma cells decreased significantly (P <0.05) with EdU staining; The inhibition of TWIST1 increased Bax expression in glioma cells, and inhibited Bcl-2 expression (P < 0.05) with Western blot; TUNEL staining further confirmed that the apoptosis level of glioma cells in the si-TWIST1 group was higher, compared with the control group (P <0.05). Finally, we found that mTOR protein expression in glioma was higher, compared with adjacent tissues. in vitro experiments showed that mTOR expression in glioma cells was decreased after the inhibition of TWIST1 (P <0.05). TWIST1 expression level in glioma was increased. The inhibition of TWIST1 inhibits the proliferation of glioma by blocking the mTOR signal pathway, and promote the apoptosis of glioma.

Erinacerins, Novel Glioma Inhibitors from Hericium erinaceus, Induce Apoptosis of U87 Cells through Bax/Capase-2 Pathway

Background: Glioma is the most common tumor of the central nervous system. Hericium erinaceus, which has been reported to have a variety of pharmacological activities, is a widely used Traditional Chinese Medicine (TCM), and also a kind of delicious food accepted by the public. Methods and Results: In this study, two new natural products, compounds 1 and 2, were isolated and identified from Hericium erinaceus. They were named erinacerin O and erinacerin P, respectively, after the structural identification, and their effects on human glioma cell line U87 were evaluated. Erinacerin P (2) exhibited obvious cytotoxicity on human glioma cell line U87. The IC50 value of 2 was 19.32μg/mL. The results showed that the apoptosis of U87 cells treated with 2 increased and the morphology of U87 cells altered significantly. Flow cytometry experiment showed that 2 could significantly increase the apoptosis rate of U87 cells and reduce DNA replication. Western blot results suggested the Bax/capase-3 pathway was involved in the U87 cell apoptosis induced by 2. Conclusion: Erinacerin O and Erinacerin P are novel compounds obtained from Hericium erinaceus and Erinacerin P could be a potential novel glioma inhibitor.

Upregulation of miRNA-202 promotes apoptosis and inhibits migration of glioma cell line via downregulation of ROCK1 gene

Background: We evaluated role(s) of miR-202 in glioma cell lines, its effect on ROCK1 expression, and also evaluation of apoptosis and migration of human glioma cell line after transfection with miR-202 mimics and inhibitors. Material and methods: The cell lines were transfected with mimic, inhibitor and NC of miR-202. Reverse transcription polymerase chain reaction (RT-PCR) was conducted to evaluate the expression of miR‐202 and ROCK1 . Western blot was performed to detect the protein level of ROCK1. Furthermore, MTT and wound healing assay were performed to evaluate the effects of miR-202 on apoptosis and migration of human glioma cell line, respectively. Results: miR-202 showed a significantly decrease in human glioma cell lines, compared with the NHA cell line (P<0.05). The ROCK1 expression was significantly upregulated in glioma cell lines, compared with the NHA cell line ( P <0.05). Furthermore, a negative correlation was observed between expression of ROCK1 and miR-202 ( P =0.01, r=-0.426). The mRNA and protein levels of ROCK1 were decreased in U87 cell line in miR-202 mimics group, compared with mimic NC group (P<0.05). In addition, apoptosis was significantly increased in miR-202 mimics, compared with the NC group in U87 cell line at 72 and 96 h (P<0.05). Furthermore, invasion showed a significant decrease in miR-202 mimic group, compared with U87 cell line at 24 and 48 h (P<0.05). Conclusions: The miR-202 could serve as a tumour-suppressor miRNA in glioma. Therefore, targeting ROCK1 by miR-202 may increase improve disease outcome and could be considered as a potential therapeutic target for glioma patients.