Antitumor mechanism of kangliu pill on gliomas in mice through PI3K-Akt signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Gliomas are common malignant intracranial tumors that have worse prognosis and pose a serious threat to human health. The Kangliu pill (KLP) is an innovative herbal compound from Xuanwu Hospital of Capital Medical University that has been clinically used for the treatment of gliomas for more than 40 years, and is one of the few drugs for primary treatment of this disorder. But the fundamental molecular mechanisms and pathways of KLP are not clear. AIM OF THE STUDY: To investigate the therapeutic mechanism of KLP in the treatment of gliomas. MATERIALS AND METHODS: An in situ xenograft model of red fluorescent protein-labeled human glioma cell line (U87-RFP) in BALB/c-nu mouse was established, and the therapeutic effect of KLP on gliomas was assessed by tumor weights and fluorescence areas. A quantitative proteomics approach using tandem mass tags combined with liquid chromatography-tandem mass spectrometry was performed to explore differentially expressed proteins (DEPs) in glioma tissues, and bioinformatics analyses including Gene Ontology analysis, pathway analysis, and network analysis were performed to analyze the proteins involved in the network therapeutic mechanisms responsible for key metabolic pathways. Cytological experiments corroborated the above analysis results. RESULTS: Network pharmacology approach screened 246 bioactive compounds contained in KLP, targeting 724 proteins and 173 potential targets of KLP for glioma treatment. The important targets obtained after visualizing the PPI network were AKT1, INS, GAPDH, SRC, TP53, etc. The KEGG enrichment results showed that 9 proteins were related to cancer, including Pathways in cancer, PI3K/AKT signaling pathway, etc. KLP had antitumor activity in gliomas, which reduced tumor weights and fluorescence areas. A number of DEPs possibly associated with gliomas were identified through quantitative proteomic techniques. Among these DEPs, 50 (25 upregulated and 25 downregulated) were identified that might be associated with KLP action. Bioinformatics showed that these 50 DEPs were mainly focused on focal adhesion, extracellular matrix (ECM)-receptor interactions, and the PI3K-Akt signaling pathway. Cytological experiments revealed that KLP significantly inhibited the proliferation and promoted apoptosis of U87-MG human glioma cells, and its mechanism was through the inhibition of PI3K/AKT signaling pathway. CONCLUSION: Therapeutic effect of KLP was regulation of multiple pathways in the treatment of gliomas. In specific, it interacts through the PI3K-Akt signaling pathway. This work may contribute proteomic insights for further research on the medical treatment of glioma using KLP.

Partial response to Chinese patent medicine Kangliu pill for adult glioblastoma: A case report and review of the literature.

BACKGROUND: Glioblastoma is the most common type of brain tumor and is invariably fatal, with a mean survival time of 8-15 mo for recently diagnosed tumors, and a 5-year survival rate of only 7.2%. The standard treatment for newly diagnosed glioblastoma includes surgery followed by concurrent chemoradiotherapy and further adjuvant temozolomide. However, the prognosis remains poor and long-term survival is rare. This report aimed to demonstrate a new therapeutic strategy for the treatment of glioblastoma. CASE SUMMARY: A patient was referred to the Department of Neurosurgery with an intracranial space-occupying lesion with a maximum diameter of approximately 5 cm. The tumor was compressing functional areas, and the patient accordingly underwent partial resection and concurrent chemoradiotherapy. The imaging and pathological findings were consistent with a diagnosis of glioblastoma with oligodendroglioma differentiation (World Health Organization IV). The patient was finally diagnosed with glioblastoma. However, the patient discontinued treatment due to intolerable side effects, and was prescribed Kangliu pill (KLP) 7.5 g three times/d, which he has continued to date. Significant shrinkage of the tumor (maximum diameter reduced from about 3.5 to about 2 cm) was found after 3 mo of KLP therapy, and the tumor was further reduced to about 1 cm after 3 years. The patient's symptoms of headache, limb weakness, and left hemiplegia were relieved, with no side effects. CONCLUSION: KLP has been a successful intervention for glioblastoma, and the current case indicates that traditional Chinese medicine may offer effective alternative therapies for glioblastoma.

[Role of CyclinD1/IgH Detection by FISH in Differential Diagnostic Significance between Mantle Cell Lymphoma and Chronic Lymphocytic Leukemia].

OBJECTIVE: To investigate the feasibility of CyclinD1/IgH detection by FISH in diferential diagnosis between mantle cell lymphoma (MCL) and chronic lymphocytic leukeamia (CLL). METHODS: The FISH detection was performed for CyclinD1/IgH fusion gene. A comprehensive analysis was carried out for clinical features, such as age, sex , WBC count and lymphocyte count, the bone marrow morphology and immunohistochemical staining were carried for CyclinD1/IgH. RESULTS: It is often difficult to distinguish MCL from CLL by bone marrow morphology, when the cell morphology was not typical; there was no difference in age, sex, WBC count and lymphocyte count between MCL and CLL groups; 9 out of 52 patients were diagnosed as MCL, and the direction of CyclinD1/IgH by FISH was positive in 7 of 9 MCL, while 3 of the 7 patients were negative by immunohistochemical staining for CyclinD1. CONCLUSION: Detection of CyclinD1/IgH by FISH can be used as a specific and feasible method for differential diagnosis of mantle cell lymphoma from chronic lymphocytic leukeamia.

[Leukemia SH-1 cells purged by ZnPcH(1)-based photodynamic therapy].

The objective of this study was to investigate the effect of a novel Zinc phthalocyanine (ZnPcH(1)) based photodynamic therapy (PDT) on acute monocytic leukemia cell lines SHI-1 and its mechanism, so as to provide theory basis for bone marrow purging in vitro for patients with leukemia. The killing effect of ZnPcH(1)-PDT on SHI-1 cells were assessed by MTT method; the SHI-1 cell death patterns were analyzed by AO/EB fluorescence staining, TdT-mediated dUTP nick end labeling (TUNEL), DNA ploidy analysis, and Annexin V-FITC/PI double staining.Cell mixture was established by integrating SHI-1 cells with normal bone marrow MNC (by 1:100-1:10 000). Purging effect of ZnPcH(1)-PDT against SHI-1 mixed into normal MNC was assessed by analyzing the expression of fusion gene MLL/AF6 mRNA using nested RT-PCR. The results showed that ZnPcH(1)-PDT could effectively inhibit SHI-1 cell proliferation in dose-dependent manner, and ZnPcH(1)-PDT could induce cell apoptosis in time-dependent manner. 0.5 µmol/L ZnPcH(1)-PDT could completely photoinactivated kill SHI-1 cells in the simulated remission bone marrow. It concluded that ZnPcH(1)-PDT may be a effective and convenient promising purging technique for leukemia.

[Apoptosis in nasopharyngeal carcinoma cell line NCE induced by curcumin and its molecular mechanism].

OBJECTIVE: To investigate the mechanism underlying the curcumin-induced apoptosis of nasopharyngeal carcinoma (NPC) cell line NCE cells. METHODS: The characteristics of apoptosis were identified by observation acridine orange and ethidium bromide stains, ultrastructure assay, DNA fragmentation assay and TdT-mediated dUTP nick end labeling method (TUNEL). Mitochondrial membrane potential (delta psi m), activity of caspase-3, cytosol cytochrome C and expression of gene Fas were determined by flow cytometry (FCM), Western Blot and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Several evidences of apoptosis were obtained from curcumin-treated NCE cells by acridine orange and ethidium bromide stains, ultrastructure identification, DNA fragmentation assay and TUNEL staining. And the mean TUNEL-positive rates increased significantly at the 3 different time points (12 h, 24 h and 48 h; 25.6%, 40.3% and 54.5%, respectively). In the curcumin-treated-groups, delta psi m altered significantly and the positive rates increased in a time-dependent manner. At the 3 different time points, the mean positive rates were 26.8%, 42.3% and 68.2%, respectively. When caspase-3 activity was detected, 80.5% cells presented proteases activities after 12 h incubation with curcumin. Western Blot analysis showed that cytoplasmic cytochrome C increased significantly after incubation with curcumin. Flow cytometry and RT-PCR analysis showed that curcumin could up-regulate the Fas expression in time-depended manner , the positive rates of Fas protein increased from 33.6% to 89.9%. CONCLUSIONS: Curcumin induced apoptosis of NCE cells both through mitochondria-dependent pathway and death receptor pathway.