Network Pharmacology to Uncover the Molecular Mechanisms of Action of LeiGongTeng for the Treatment of Nasopharyngeal Carcinoma.

BACKGROUND Nasopharyngeal carcinoma (NPC) is a common head and neck cancer epidemic in southern China and southeast Asia. LeiGongTeng has been widely used for the treatment of cancers. The purpose of this study was to determine the pharmacological mechanism of action of LeiGongTeng in the treatment of NPC using a network pharmacological approach. MATERIAL AND METHODS The traditional Chinese medicine systems pharmacology (TCMSP) database was used to identify active ingredients and associated target proteins for LeiGongTeng. Cytoscape was utilized to create a drug-disease network and topology analysis was conducted to analyze the degree of each ingredient. The Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) online tool was applied for the construction and analysis of the protein-protein interaction (PPI) network, while Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment and Gene Ontology (GO) functional analyses were utilized to determine drug-disease common genes. RESULTS 22 active ingredients including kaempferol, nobiletin, and beta-sitosterol, and 30 drug-disease common genes including VEGFA, CASP3, ESR1, and RELA were identified. GO analysis indicated that 94 biological processes, including RNA polymerase II, apoptotic process, response to drug, cell adhesion, and response to hypoxia, were found to be associated with NPC. The KEGG enrichment analysis showed that 58 pathways, including the PI3K-Akt signaling pathway, microRNAs in cancer, tumor necrosis factor (TNF) signaling pathway and pathways in cancer were found to be associated with NPC. CONCLUSIONS LeiGongTeng exerts its therapeutic effect through various biological processes and signaling pathways since it acts on several target genes. Systematic pharmacology can be used to predict the underlying function of LeiGongTeng and its mechanism of action in NPC.

Small molecule inhibitor TW-37 is tolerable and synergistic with chemotherapy in nasopharyngeal carcinoma.

Chemotherapy is a crucial adjuvant therapy of advanced nasopharyngeal carcinoma (NPC). However, enhancing sensitivity and tolerance of chemotherapeutics in NPC treatment have been challenging. Both Bcl-2 and Mcl-1, 2 pro-survival proteins of Bcl-2 family, play essential roles on the chemotherapy tolerance of numerous cancers. In the present study, we explored the influences of TW-37, a small molecule inhibitor of Bcl-2 and Mcl-1, on the efficiency of chemotherapy for NPC. Oncomine cancer database shows that NPC tissues have higher expression of Bcl-2 and Mcl-1 than those of normal nasopharyngeal epithelial (NPE) tissues. And our results reveal that chemotherapeutics, Cisplatin (CDDP) and 5-Fluoracil (5-FU), result in the greater decrease of protein level of Bcl-2 and Mcl-1 in NPC cells than those in NPE cells. TW-37 does not have significant impact on the chemotherapeutics-treated NPE cell viability at a dosage that efficiently reduces chemotherapeutics-treated NPC cell viability. Moreover, impacts of TW-37 on the cell viability of chemotherapeutics-treated NPC cells are dependent on the expression of Bcl-2 and Mcl-1 in NPC cells. Further explorations suggest that TW-37 prominently promotes apoptosis in NPC cells under chemotherapeutics treatments but not in NPE cells. Meanwhile, TW-37 also remarkably reduces colony formation ability of chemotherapeutics-treated NPC cells. Importantly, in vivo models, TW-37 observably increases chemosensitivity of NPC tumors but has not markedly influence on the normal tissues in mice. In conclusion, our results point to TW-37 as a promising ancillary drug for the chemotherapy of NPC.

[Research on the effect of biyan qingdu granula drug-containing serum on inhibition of nasopharyngeal carcinoma cells growth in vitro].

OBJECTIVE: To investigate the effect of Biyan Qingdu Granula drug-containing serum (BQG-DS) on cell growth and apoptosis in nasopharyngeal carcinoma cell lines CNE1, CNE2, TWO3, C666-1, and explore the antineoplastic mechanism of Biyan Qingdu Granula. METHODS: SD rats were randomly divided into three groups: experimental (Biyan Qingdu Granula) group, positive control (cytoxan) group and negative control group. After administration of drug, the serum was collected from the treated animals. MTT assay was used to examine the effect of BQG-DS on the proliferation of CNE1, CNE2, TWO3, C666-1 cell, and flow cytometry was used to observe the cell cycle distribution. Apoptosis of CNE1, CNE2, TWO3, C666-1 cell was further investigated by inverted microscope. RESULTS: BQG-DS inhibited the proliferation of CNE1, CNE2, TWO3, C666-1 cell and the effects were in a time-and concentration-dependent manner. BQG-DS could also induce apoptosis while the G1 phase was arrested. CONCLUSION: BQG-DS inhibits proliferation of nasopharyngeal carcinoma cells via induction of apoptosis and arrest of cell cycle.

Long-term efficacy of microwave hyperthermia combined with chemoradiotherapy in treatment of nasopharyngeal carcinoma with cervical lymph node metastases.

OBJECTIVE: The long-term efficacy of microwave hyperthermia combined with chemoradiotherapy in treating nasopharyngeal carcinoma (NPC) with metastatic foci in cervical lymph nodes was evaluated. METHODS: A total of 154 cases of N2 or N3 stage NPC were randomized into two groups: hyperthermia group (76 cases) and control group (78 cases). Both received cisplatin chemotherapy and radiotherapy. In addition, the hyperthermia group further received microwave hyperthermia to the metastatic cervical nodes with different patterns (before or after radiotherapy), heating temperatures (T90 < 43° and T90 ≥ 43°) and hyperthermia episodes (< 4 times, 4-10 times and > 10 times). RESULTS: The 3-month and 5-year complete response (CR) rates of cervical lymph nodes in the hyperthermia group were significantly higher than those in the control group. The 5-year disease-free survival (DFS) rate and the 3-year / 5-year overall survival rate in the hyperthermia group were also significantly higher. There was no significant difference in 5-year metastatic rates. In the hyperthermia group, the 3-month and 5-year CR rates of T90 < 43° treatment were significantly lower than with T90 ≥ 43° treatment. The CR rate was highest when the hyperthermia was performed 4-10 times. There were no significant differences in 3-month and 5-year CR rates between hyperthermia before or after radiotherapy treatment. CONCLUSION: Microwave hyperthermia combined with chemoradiotherapy can increase local control, DFS and 3, 5-year overall survival rates of patients with N2 ~ N3 stage NPC. The heating temperature should be over 43° with hyperthermia repeated 4-10 times.

Tea polyphenols protect against irradiation-induced injury in submandibular glands' cells: a preliminary study.

AIM: To study the protective effect of tea polyphenols (TPs) on submandibular glands affected by radiation injury. METHODS: Sixty rats were randomly divided into radiation group (R-group, N = 30) and TP-pre-treated-radiation group (TPR-group, N = 30). The rats were intragastrically administered with TP or normal sodium from 14 days before radiation, continuously daily, until the experiment. All the rats in both groups were irradiated with a single exposure dose of 15 Gy gamma rays that were delivered to the head and neck areas. Ten rats of each group were anatomised on the 3rd, 6th and 30th day after irradiation, respectively. The submandibular glands of the rats were removed for the study. The morphologic changes of the submandibular glands were observed by transmission electron microscopy (TEM). The terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labelling (TUNEL) method was used to detect apoptosis of the submandibular glands' cells. RESULTS: Electron microscope observation of the submandibular glands showed that the lesions of the TPR-group were mild. Change in apoptosis of the cells was not obvious compared with the R-group. The cell apotosis was typical after irradiation in the R-group. Apoptosis index that was detected in the cells of submandibular glands of the TPR-group was statistically significantly decreased compared with the R-group (P < 0.01) on the 3rd, 6th and 30th day after irradiation. CONCLUSION: TP could protect submandibular glands from radiation injuries, and the protection mechanism may be realised by anti-apoptosis.

[Scavenging effect of Sarcandra glabra powder on radiation-induced reactive oxygen species in the parotid gland of miniature pigs].

OBJECTIVE: To investigate the effect of Sarcandra glabra in scavenging reactive oxygen species (ROS) produced by γ-ray irradiation in the parotid gland of miniature pigs. METHODS: Forty-five male miniature pigs were randomly divided into control group, radiation group and radiation plus medication group, and each group contained 3 parallel groups (subgroups a, b and c). From 1 week before exposure of the parotid gland region to 15 Gy γ-ray irradiation (which was not administered in the control group), the miniature pigs in radiation plus medication group were given Sarcandra glabra powder, while those in the other groups received an equal amount of saline. Bilateral parotid glands were taken and weighed on the days 10, 40 and 90 following the exposure in subgroups a, b, and c, respectively, and ROS content in the parotid glands were determined by enzyme-linked immunosorbent assay. RESULTS: The content of ROS was significantly lower in radiation plus medication group than in the radiation group (P<0.01). In the radiation plus medication group, the ROS content showed no significant difference between subgroups a and b or between subgroups a and c (P>0.01), but differed significantly between subgroups b and c (P<0.01). Sarcandra glabra showed a strong ROS-scavenging effect 10 days after the irradiation, and the ROS content was similar with that in the control group (P>0.01); at 40 and 90 days, the ROS-scavenging effect of Sarcandra glabra was still observable, but the ROS content was significantly higher in the irradiation plus medication group than in the control group (P<0.01). CONCLUSION: Sarcandra glabra displays a ROS-scavenging effect in the parotid gland of miniature pigs against irradiation, especially at 10 days following the exposure, which may serve as the main mechanism for the protective effect of Sarcandra glabra against radiation injury in the parotid gland.