Aim To investigate the effects of PLCε on the invasion and migration of human osteosarcoma cancer cells U2OS. Methods RNA interference ( RNAi) was used to inhibit PLCεexpression, and the proliferation of cancer cells was measured by CCK-8 assay. The migration of the cells was measured by scratch wound healing assay and migration chamber as-say. Gelatin zymography was performed to measure the MMP2 activities in U2OS cells. Results PLCε ex-pression was suppressed by siRNA. CCK-8 assay showed that PLCε had no effect on the proliferation of cancer cells. PLCε knockdown inhibited cell invasion and activities of MMP2 . Conclusion PLCε knock-down can inhibit the migration and invasion of human osteosarcoma cancer cells U2OS.

We report a case of cushing's syndrome caused by ectopic adreocortical adenoma. The patient is a 37 years old woman, she was admitted to our hospital for " 2 years history of hypertension and weakness in both lower extremities for 2 months". Physical examination revealed: blood pressure 160/116 mmHg(1 mmHg＝0.133 kPa), body mass index 27.47 kg/m2, moon-face, increased fat in the neck and back, purple marks on abdominal skin, withⅡdegree edema of both lower extremities. Laboratory examination revealed that serum cortisol levels were elevated, loss of normal circadian rhythm, and serum adrenocorticotropic hormone (ACTH) was suppressed, the level of cortisol could not be suppressed in low dose desamethasone suppression test. Adrenal computed tomography ( CT) revealed a nodule in the right retroperitoneum, compression of the renal hilum, no bilateral adrenal adenoma and hyperplasia were found. This patient was diagnosed as corticotropin-independent Cushing's syndrome unequivocally. The clinical symptoms were relieved after successful laparoscopic retroperitoneum resection of the nodule. Pathological exam confirmed adrenocortical adenoma in ectopic adrenal tissue. Thus, we should consider the ectopic corticosteroid-secreting tumor in the context of corticotropin-independent Cushing's syndrome, especially when the imaging studies of adrenal revealed bilateral adrenal glands were normal or atrophic, which helped to make an appropriate strategy treatment.

Background and objective Rap2a, a member of the small GTPase superfamily, plays a critical role in regulating the function of integrin and cell adhesion, thereby controlling cell motility and cell/matrix interactions. However, the function of Rap2a in carcinogenesis is still poorly understood. To clone Rap2a cDNA, which belongs to human Ras-related small G protein superfamily, we constructed its eukaryotic expression vector and determined its expression in lung cancer cells. hTe aim of this study is to explore the role of Rap2a in carcinogenesis. Methods hTe levels of endogenous Rap2a protein in lung cancer cells were measured by Western blot. Total RNA of human osteosarcoma cells U2OS was extracted and reverse-transcribed into cDNA by RT-PCR. hTen, Rap2a gene was ampliifed by PCR and inserted into pcDNA3.1(+). hTe re-constructed plasmid was identiifed by restricted enzyme digestion and sequencing. pcDNA3.1(+)-Rap2a was transfected into H1299 and A549 cells, the expression of Rap2a was detected by Western blot. In addition, the migratory abilities of lung cancer cells were evaluated by Transwell assay. Matrix metalloproteinase (MMP)2 enzyme activity was evaluated by gelatin zymogra-phy. Results Rap2a is signiifcantly upregulated in lung cancer cells. hTe results of enzyme digestion and sequencing showed that the coding sequence of pcDNA3.1(+)-Rap2a was right and was inserted into the vector correctly. hTe results of Western blot showed that H1299 and A549 cells were transfected successfully. Transwell assay indicated that the ectopic expression of Rap2a promotes lung cancer cells migration. Correspondly, enzyme activity of MMP2 also increased. Conclusion Eukaryotic expression plasmid pcDNA3.1(+)-Rap2a was constructed successfully. Rap2a could be expressed in lung cancer cells effciently and promotes lung cancer cell migration.