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OBJECTIVE@#To explore the influences of andrographolide (Andro) on bladder cancer cell lines and a tumor xenograft mouse model bearing 5637 cells.@*METHODS@#For in vitro experiments, T24 cells were stimulated with Andro (0-40 µmol/L) and 5637 cells were stimulated with Andro (0 to 80 µmol/L). Cell growth, migration, and infiltration were assessed using cell counting kit-8, colony formation, wound healing, and transwell assays. Apoptosis rate was examined using flow cytometry. In in vivo study, the antitumor effect of Andro (10 mg/kg) was evaluated by 5637 tumor-bearing mice, and levels of nuclear factor κ B (NF- κ B) and phosphoinositide 3-kinase/AKT related-proteins were determined by immunoblotting.@*RESULTS@#Andro suppressed growth, migration, and infiltraion of bladder cancer cells (P⩽0.05 or P⩽0.01). Additionally, Andro induced intrinsic mitochondria-dependent apoptosis in bladder cancer cell lines. Furthermore, Andro inhibited bladder cancer growth in mice (P⩽0.01). The expression of p65, p-AKT were suppressed by Andro treatment in vitro and in vivo (P⩽0.05 or P⩽0.01).@*CONCLUSIONS@#Andrographolide inhibits proliferation and promotes apoptosis in bladder cancer cells by interfering with NF- κ B and PI3K/AKT signaling in vitro and in vivo.
Subject(s)
Animals , Humans , Mice , Apoptosis , Cell Line, Tumor , Cell Proliferation , Diterpenes/therapeutic use , NF-kappa B/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Urinary Bladder Neoplasms/drug therapyABSTRACT
Objective:Dorsocervical fat pad is common in middle-aged women. Current treatments include surgical excision and liposuction. We evaluated the therapeutic effect of tumescent liposuction on dorsocervical fat pad. Anatomical study was also carried out to explore the anatomical structure and significance of dorsocervical fat pad.Methods:From Jan. 2009 to Dec. 2018, twenty-seven patients with dorsocervical fat pad were treated with tumescent liposuction in Peking University Third Hospital. Small incisions were made in bilateral scapular region and 4 mm suction cannula was applied. A female cadaver fixed with formaldehyde was dissected to investigate the structure of posterior cervical and dorsal region. The specimens were stained with HE and Masson staining.Results:14 patients were followed up for no less than 6 months, with an average follow-up time of 27 months. Patients＇ dorsocervical area were flat and smooth after the surgery. Patient satisfaction rate was 100% and no severe complication was reported except bruise and pain. The symptoms of dorsocervical pain in two patients were significantly improved after operation. Anatomical study showed that the dorsocervical fat pad was composed of superficial and deep layer of adipose tissue, with clear boundary between the two layers and no obvious capsule. The collagen fibers in deep layer were more and denser than those in superficial layer.Conclusions:Tumescent liposuction can effectively treat dorsocervical fat pad. The surgery outcome is ideal with little complication.Through the study of the anatomical structure of the dorsocervical fat pad, the operation method and principle of liposuction can be improved and the operation efficiency can be enhanced.
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Primary lesion removal and lymph node dissection are the main constituents of radical gastrectomy. However, the high recurrence rate after D2 radical gastrectomy for advanced gastric cancer has not improved. Recently, studies have found that discrete tumor deposits in the mesogastrium may be an important factor affecting the prognosis of gastric cancer after surgery. With the development of laparoscopic equipment, the ever-expanding "submicroscopic vision" makes it possible to completely remove the mesogastrium. Professor Gong Jianping advocated "membrane anatomy" to optimize the concept of radical gastrectomy: D2- based complete mesenteric resection (CME), namely D2+CME procedure. To prevent the leakage of tumor cells into the surgical field, as histological barrier, the intact mesogastrium should be located. The essential difference between D2+CME and previous D2/D2+systematic mesogastrium excision (SME), en-bloc mesogastric excision (EME) is as follow: double-factor guiding (lymph nodes and discrete tumor deposits) vs. single factor guiding (lymph nodes only). After practicing dozens of radical gastrectomy (D2+CME) authors believe that its conceptual connotation (double factor guiding) and operational extension (above mesentery bed) cover D2. In D2+CME surgery, depending on the anatomical identification under the magnified field of view, the conformal space between gastric mesentery and mesenteric beds is unique operational plane with repeatability. These findings and considerations address one problem: where is the precise boundary of en bloc principle in radical gastrectomy? In author′s opinion, with laparoscopy and "sub-microsurgery" progression and detection of discrete tumor deposit metastasis, survival benefit from definition of en bloc boundary in radical gastrectomy will be widely recognized. Meanwhile, D2+CME procedure is an appropriate way for study. Although the development of the "membrane anatomy" concept for gastric cancer still requires many further clinical and basic researches, it is reasonable to foresee that D2+CME surgery will guide a concept-optimized era for gastric cancer surgery.
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Objective:To study the inhibitory effects of andrographolide (Andro) on the proliferation,migration and clone formation ability of renal cell carcinoma (RCC) cells and the induction on the apoptosis,and to clarify their related mechanisms.Methods:The RCC cells were treated with different concentrations (5,10,20,40 and 80 μmol · L-1) of Andro as experimental groups,and 0μmol · L 1 Andro group was used as blank control group,MTS assay was used to detect the proliferation rates of RCC cells in various groups.The RCC cells were treated with different concentrations (0.50,1.25 and 2.50 μmol · L-1) of Andro as experimental groups,and 0 μmol · L 1Andro group was used as blank control group.Clonogenic assays was used to detect the colony formation ability of RCC cells in various groups.The RCC cells were treated with different concentrations (10,20and 40 μmol · L-1) of Andro as experimental groups,and 0 μmol · L-1 Andro group was used as blank control group.Wound healing assay was used to detect the migration ability of RCC cells in various groups.Flow cytometry was used to detect the apoptotic rates of RCC cells in various groups.Western blotting was performed to detect the expression levels of apoptosis related proteins in RCC cells in various groups.Results:Compared with blank control group,the proliferation rates of RCC cells in 10,20,40 and 80 μmol · L-1 Andro groups were markedly decreased (P＜0.05 or P＜0.01).Compared with blank control group,the colony formation rates of RCC cells in 0.50 and 1.25μmol · L-1 Andro groups were markedly decreased (P＜0.05 or P＜0.01).Compared with blank control group,the scratch healing rates of RCC cells in 10,20 and 40 μmol · L-1 Andro groups were markedly decreased (P＜0.01),and the apoptotic rates of RCC cells in 20 and 40 μmol · L-1 Andro groups were markedly increased (P＜0.01).Compared with blank control group,the expression level of γ-H2AX protein in 40μmol · L-1 Andro group was markedly increased (P＜0.01),the expression level of Caspase-8 protein was decreased (P＜0.05),and the expression level of cleaved Caspase-8 protein was markedly increased (P＜0.01).Conclusion:Andro can effectively suppress the proliferation,migration and colony formation ability of RCC cells and induce the apoptosis of RCC cells.The mechanism of apoptosis might be related to inducing the DNA damage and the apoptotic pathways induced by JNK / H2AX and Caspase-8.
ABSTRACT
Objective: To study the inhibitory effects of andrographolide (Andro) on the proliferation, migration and clone formation ability of renal cell carcinoma (RCC) cells and the induction on the apoptosis, and to clarify their related mechanisms. Methods: The RCC cells were treated with different concentrations (5, 10, 20, 40 and 80 μmol · L-1) of Andro as experimental groups, and 0 μmol · L-1 Andro group was used as blank control group, MTS assay was used to detect the proliferation rates of RCC cells in various groups. The RCC cells were treated with different concentrations (0. 50, 1. 25 and 2. 50 μmol · L -1) of Andro as experimental groups, and 0 jumol · L-1 Andro group was used as blank control group. Clonogenic assays was used to detect the colony formation ability of RCC cells in various groups. The RCC cells were treated with different concentrations (10, 20 and 40 μmol · L-1) of Andro as experimental groups, and 0 μmol · L-1 Andro group was used as blank control group. Wound healing assay was used to detect the migration ability of RCC cells in various groups. Flow cytometry was used to detect the apoptotic rates of RCC cells in various groups. Western blotting was performed to detect the expression levels of apoptosis related proteins in RCC cells in various groups. Results: Compared with blank control group, the proliferation rates of RCC cells in 10, 20, 40 and 80 μmol · L-1 Andro groups were markedly decreased (P<0.05 or P<0.01). Compared with blank control group, the colony formation rates of RCC cells in 0. 50 and 1. 25 jumol · L-1 Andro groups were markedly decreased (P<0. 05 or P<0. 01). Compared with blank control group, the scratch healing rates of RCC cells in 10, 20 and 40 μmol · L-1 Andro groups were markedly decreased (P<0. 01), and the apoptotic rates of RCC cells in 20 and 40 μmol · L-1 Andro groups were markedly increased (P<0. 01). Compared with blank control group, the expression level of γ-H2AX protein in 40 jumol · L-1 Andro group was markedly increased (P<0. 01), the expression level of Caspase-8 protein was decreased (P<0. 05), and the expression level of cleaved Caspase-8 protein was markedly increased (P<0. 01). Conclusion: Andro can effectively suppress the proliferation, migration and colony formation ability of RCC cells and induce the apoptosis of RCC cells. The mechanism of apoptosis might be related to inducing the DNA damage and the apoptotic pathways induced by JNK/H2AX and Caspase-8.