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Cancer is one of the major diseases that threaten human health, and to find effective anticancer drugs has become an important research topic at home and abroad. Camptothecin (camptothecin, CPT) is an alkaloid extracted from Chinese Davidia involucrata plants of C. acuminata in, with a strong anti-tumor activity, can be used for the treatment of several types of cancer, such as stomach cancer, liver cancer, bladder cancer and leukemia. But there are also bone marrow suppression, vomiting, diarrhea and severe hemorrhagic cystitis side effects, as well as poor water solubility, bioavailability and low defects. DNA topoisomerase Ⅰ (Topoosomerase Ⅰ, Topo Ⅰ) is the main target of the CPT and its analogues, anti-tumor effect by copying the conflict model to explain. The subject combination of in vivo and in vitro experiments, the screening of new camptothecin derivative (Camptothecin the derivates, CD), and by observing the immunological parameters, apoptosis, cell cycle and multi-drug resistance, preliminary study mechanism of action of the derivative to provide the experimental basis for the development of new drug for anti-tumor. The main contents are as follows: 1.CD screening 1.1.CD of in vitro screening CD-Ⅶ, Ⅷ drug concentration for 10 -4 ~ 10 -8 MoL / L dose under HepG2, L1210, SMMC-7721 cell lines was inhibited, was a certain dose, time-effect relationship. CD-I, II, III, IV, V, VI, IX, X, no significant inhibitory effect on tumor cells, CD-XI, XII SMMC-7721 and L1210 tumor cell lines also have some effect. Vivo screening in 1.2.CD CD-VIII significantly inhibited the growth of S180 tumor bearing mice tumors with higher tumor inhibition rate and increases with the concentration, the inhibition was enhanced. CD-VII S180 tumor bearing mice significantly inhibited. Tip: CD-VIII may have strong anti-tumor activity. 2.CD-VIII preliminary study on anti-tumor mechanism of action allows the tumor-bearing mice with low 2.1.CD-Ⅷ part of the immune function of tumor-bearing mice CD-Ⅷ 3.0,1.5 mg / kg ConA-induced lymphocyte proliferative response enhancement, 0.75, 1.5, mg / kg dose also low LPS-induced lymphocyte proliferation was enhanced, in addition to the tumor-bearing mice, but also increased the tumor-bearing mice spleen index, thymus index, can significantly reduce the elevated NO levels of tumor-bearing mice, tumor-bearing mice TNF-alpha and IL-2 level, and was dose-dependent. Tip: CD-VIII anti-tumor effect may be associated with enhanced tumor-bearing mice immune function. The 2.2.CD-VIII-induced apoptosis of tumor cells and mechanisms using staining Hochest33258, agarose gel electrophoresis and flow cytometry to detect CD-VIII the different doses role HepG2 cells, the situation after the results appear typical apoptotic features, such as \CD-VIII can make GRP78 mRNA expression increased significantly, while the CPT group had no significant effect;, P-JNK expression significantly increased activation of JNK earlier than apoptosis. Tip: CD-VIII-induced apoptosis may be associated with the endoplasmic reticulum stress response pathway of apoptosis. 2.4.CD-VIII LLCPK / CMV and LLCPK/Mrp2 of cell lines resistant tumor cell lines for the study, found that the DDP and CPT resistance, resistance index were 2.705 and 2.963 times, there was a significant difference, while the CD-VIII, 5-FU is more sensitive. At the same time, the CD-VIII group LLCPK/Mrp2 cells MRP2 mRNA expression can reduce the CPT group Mrp2 expression had no significant effect. Prompt: CD-VIII may enhance the sensitivity of drug-resistant tumor cells by inhibiting expression of LLCPK/Mrp2 cell Mrp2mRNA of.
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