Mechanism of PTPN18 for regulating the migration and invasion of endometrial cancer cells via the MYC/PI3K/AKT pathway.

OBJECTIVE: Endometrial cancer (EC) is a prevalent gynecologic malignancy. The critical role of PTPN18 in EC has been reported, while its role in the aerobic glycolysis of EC cells remains unclear. Our current study focused on the mechanism of PTPN18 in the regulation of aerobic glycolysis in EC. METHODS: PTPN18 expression levels in endometrial stromal cells (KC02-44D) and EC cells (KLE, HEC-1-A, HEC-1B, and HEC-50) were determined. Following transfection of sh-PTPN18 in HEC-1-A cells, the changes in cell migratory and invasive abilities were assessed by the Transwell assay, and the changes in glucose consumption, lactic acid secretion, and ATP levels were detected using kits. The expression levels of glycolysis-related proteins HIF-1α, PKM2, and LDHA and the activation of the MYC/PI3K/AKT pathway were detected by Western blot. Additionally, sh-PTPN18 and pcDNA3.1-MYC were transfected into HEC-1-A cells to further explore their roles in the changes in aerobic glycolysis, migration, and invasion ability of EC cells. RESULTS: Expression of PTPN18 in EC cells was up-regulated (HEC-1-A>HEC-1B>HEC-50>KLE). PTPN18 knockdown suppressed EC cell migration and invasion. Additionally, PTPN18 knockdown reduced glucose consumption, lactate production, ATP levels, and glycolysis-related protein levels (HIF-1α, PKM2, LDHA). PTPN18 knockdown inhibited the activation of the MYC/PI3K/AKT pathway in EC cells. MYC overexpression partially annulled the inhibitory effects of PTPN18 knockdown on aerobic glycolysis, migration, and invasion of EC cells. CONCLUSION: Our present study provided evidence that the knockdown of PTPN18 inhibited the aerobic glycolysis, migration, and invasion of EC cells by suppressing the MYC/PI3K/AKT pathway.

Establishment of an animal model of adjacent segment degeneration after interbody fusion and related experimental studies.

BACKGROUND: Degenerative spine conditions are common and frequent clinical diseases, and adjacent segment disease (ASD) after spinal fusion (SF) is a common complication after spinal fusion (SF). In this study, we established an animal model of ASD after interbody fusion to observe the morphologic changes of adjacent segment (AS) disks and to determine the expression and significance of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1ß) in ASD tissues to provide a good experimental basis and reference for clinical prevention and treatment of ASD after interbody fusion. METHODS: Thirty-six male and female New Zealand rabbits weighing 2.0-2.5 kg were randomly divided into control group (group A) and experimental groups (groups B, C, and D), with 9 rabbits in each group, of which groups B, C, and D were the 4-, 8-, and 12-week groups, respectively. Autologous iliac bone grafts were used as the bone graft material. In the experimental groups, a SF was performed on the C2-C3 intervertebral space. The C3-4 adjacent segments were examined. In the experimental group, the animals were subjected to gross observation, X-ray examination, hand touch inspection, and micro-computed tomography (micro-CT) 4, 8, and 12 weeks after surgery. The micromorphologic changes of the cervical disks in the segments of the control group and experimental groups were observed under light microscopy. Immunohistochemistry and Western blotting were used to detect the expression of TNF-α and IL-1ß in the AS tissues after interbody fusion in the control and experimental groups. RESULTS: The measurement data of the rabbit cervical spine bony structures indicated that the length of the vertebral body and the sagittal diameter of the lower end of the vertebral body decreased gradually from the 2nd-6th cervical vertebrae, and the difference was statistically significant (P < 0.05). The difference in the transverse diameter of the lower end of the vertebral body was not statistically significant (P > 0.05), the change in the oblique diameter of the lower end of the vertebral body fluctuated, and the difference was statistically significant (P < 0.05). The fusion rate of the cervical spine by hand touch inspection was 22.2% (2/9), 55.6% (5/9), and 88.9% (8/9) in groups B, C, and D, respectively. The differences in bone volume-to-total volume (BV/TV) and X-ray scores were statistically significant in groups B, C, and D (P < 0.05). Significant degeneration occurred in groups B, C, and D compared with group A. The expression of TNF-α and IL-1ß in the intervertebral disk tissue was significantly higher in groups B, C, and D compared with group A (P < 0.05), and increased with time. CONCLUSION: In this study, an animal model of ASD after interbody fusion fixation in rabbits was successfully established. Postoperative imaging and hand touch inspection showed a positive correlation between the amount of new intervertebral bone and the degree of fusion with time. The results of immunohistochemistry and Western blot showed that TNF-α and IL-1ß were highly expressed in the AS tissues of the experimental group after interbody fusion, and the degree of disk degeneration was positively correlated with the time after interbody fusion.

IGFBP­rP1 affects the proliferation, apoptosis and macrophage polarization of endometrial cancer cells by regulating the PI3K/AKT pathway.

Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) is a potential tumor suppressor gene in a variety of cancers including colorectal cancer and breast cancer. However, its role and the potential mechanism in endometrial carcinoma (EC) are still unclear. The purpose of this study was to explore the effect of IGFBP-rP1 on EC cell proliferation and apoptosis and its underlying mechanism. Western blot analysis and reverse transcription-quantitative PCR were used to assess protein and gene expression levels of IGFBP-rP1 in EC cells. Overexpression of IGFBP-rP1 and/or AKT serine/threonine kinase was used to analyze their effects on cell proliferation and apoptosis of the EC cells. Co-immunoprecipitation and glutathione S transferase pull-down assays were used to analyze the interaction between IGFBP-rP1 and AKT. The expression of IGFBP-rP1 in EC cells was downregulated. Overexpression of IGFBP-rP1 inhibited the proliferation and induced apoptosis of EC cells, which were abolished by the overexpression of AKT. In addition, IGFBP-rP1 directly interacted with AKT to inhibit PI3K/AKT signaling. Additionally, M0 macrophages were induced by EC cells to differentiate into M2 macrophages, which was reversed by IGFBP-rP1. Overexpression of AKT in EC cells abolished the inhibitory effect of IGFBP-rP1 on M2 polarization. IGFBP-rP1 as an oncogenic factor inhibits M2 polarization of TAMs through PI3K/AKT signaling pathway and may be a potentially valuable target for EC therapy.

Clinical Application of Percutaneous Transforaminal Endoscopic Surgery in Lumbar Discal Cyst.

BACKGROUND: Discal cyst is very rare and can cause intractable low back pain and radiating leg pain. Symptoms are hard to distinguish from lumbar disc herniation. The best treatment for discal cyst is controversial. Most lumbar discal cysts are treated surgically, while most studies of percutaneous transforaminal endoscopic surgery are case reports. This study investigated the clinical value of percutaneous transforaminal endoscopic surgery for lumbar discal cyst. METHODS: A retrospective study was conducted in 9 patients with a discal cyst from June 2016 to November 2018. All patients had been treated by percutaneous transforaminal endoscopic surgery via a superior vertebral pedicle notch approach. Surgical outcomes were evaluated preoperatively and postoperatively using a visual analog scale for leg pain and the Oswestry Disability Index. At the final follow-up, patients were evaluated for clinical efficacy using modified Macnab criteria. RESULTS: All 9 patients had remission of symptoms after removal of discal cysts. Postoperative magnetic resonance imaging showed that all patients had complete excision of discal cysts and complete decompression of the treated segment. There were no recurrent lesions during follow-up. Mean operative time was 68.67 ± 14.02 minutes. Mean hospitalization time was 4.22 ± 1.64 days. Preoperative visual analog scale and Oswestry Disability Index score improved significantly after surgery. Visual analog scale leg score improved from 7.88 ± 1.05 preoperatively to 1.78 ± 0.66 at final follow-up (P < 0.05), and ODI score improved from 53.65 ± 12.46 to 16.25 ± 8.76 (P < 0.05). According to the modified Macnab criteria, 5 patients (55.6%) were rated excellent, 3 patients (33.3%) were rated good, and 1 patient (11.1%) was rated fair at final follow-up, with an overall excellent and good rate of 88.9%. There were no serious complications during follow-up. CONCLUSIONS: Percutaneous transforaminal endoscopic surgery could be a safe, minimally invasive surgical treatment for discal cyst, particularly suitable for patients who cannot undergo general anesthesia.

Percutaneous Endoscopic Surgery for Lumbar Discal Cyst: Two Case Reports.

Discal cyst is a rare disease, the pathogenesis is not yet clear and its symptoms are very similar to lumbar disc herniation. Although some cases may regress spontaneously, most cases of lumbar discal cysts are treated surgically. At present, there is no consensus on the treatment of this disease. The authors report the clinical usefulness of the percutaneous endoscopic transforaminal surgery technique in two patients with the lumbar 4-5 discal cyst. The clinical symptoms of both patients were unilateral lower extremity pain and lower back pain. Magnetic resonance imaging of the lumbar spine revealed lumbar discal cysts, causing compression to the spinal dura and roots. Both patients received conservative treatment for more than 6 months, but the clinical symptoms persisted so surgical treatment by percutaneous endoscopic transforaminal surgery without additional discectomy was performed under local anesthesia. The symptoms were relieved immediately after removal of the discal cysts. Postoperative magnetic resonance imaging showed that both patients had complete excision of discal cysts and complete decompression of the treated segmental. There were no recurrent lesions and complications during the follow-up period. We believe that percutaneous transforaminal endoscopic surgery could be a safe, mini-invasive and appropriate method for the treatment of discal cysts.