| Cancer is one of commonly and frequently encountered diseases threatening the human health seriously. However,the use of adjuvant chemotherapy drugs on clinical had been brought to patients more and more side reactions. So, people pay increasing attention to the natural medicines as it has less toxicity, low side reactions than the synthesis drugs.Fructus Trichosanthis is the dry friut of Trichosanthes kilirilowii Maxim or Trichosanthes rasthornii Harms which is commonly in the treatement tumers with some anti-tumer Chinese herbal compound. To investigate the effect of the Fructus Trichosanthis Senticosus(FTS) in the proliferation and apoptosis of Hela cells in vitro, the serum containing FTS were prepared with serologic pharmaeological method,and treated the Hela cells with contained herb serum of FTS in high-middle-low concentration for 24h and 48h. The proliferation-inhibiting capability of Hela cells was investigated by MTT assay.The apoptosis effect of serum containing FTS on Hela cells were analyzed by Wright′s-Giemsa staining. DNA fragmentation was performed by agarose gel electrophoresis.The results showed that Hela cells were observed some typical morphological features and super-microstructural changes of apoptosis after treatment with different concentration serum containing FTS. The growth inhibition is in dose-dependent and time-dependent manner. The greatest inhibition rate was 48.2% observed in the treatment group of Hela cells by 20% high-dosage groups of serum containing FTS for 48h. Agarose gel electrophoresis showed characteristic"DNA ladder"pattern. This suggests that the FTS can inhibit proloferation and induce apoptosis of Hela cells in vitro.Blaps femoralis Fischer-Waldheim (BFFW) is a dominance guilds of insect community, which belongs to Blaps Fabricius of Blaptini of Tenebrionidae. This kinds of insects were often used to treatment emphysema, rheumatism, and tumor, etc. In order to identify the antitumer active component of BFFW, the serum containing BFFW were prepared with serologic pharmaeological method firstly. Then, the inhibition to cell growth and the proliferation of Hela cells were measured with the serum according to methods above. The results showed that morphological and super-microstructural changes of Hela cells treated with the serum containing BFFW were observed by inverted microscope, double staining with AO-EB. The inhibition is in dose-dependent and time-dependent manner. The greatest inhibition rate was 37.3% observed in the treatment group of Hela cells by 20% high-dosage groups of serum containing BFFW for 48h.To investigate the active anti-tumer components of BFFW, we obtained the protein extract with the ammonium sulfate precipitation. The morphological and super-microstructural changes of apoptosis were observed with inverted fluorescence microscope after Hela cells were treated by protein extract of BFFW (7.5μg/mL, 10.0μg/mL, 15.0μg/mL) for 48h in vitro. DNA ladder assay and flow cytometry (FCM) were carried out for observed the inducible apoptosis by BFFW in Hela cells. FCM measured apoptosis rates of four experimental groups (0μg/mL, 7.5μg/mL, 10.0μg/mL, 15.0μg/mL treatment for 48h or 72 h, respectively). Western blot analyzed the expression of caspase-3 of Hela cells treated with 7.5μg/mL,10.0μg/mL,15.0μg/mL for 48h or 72 h, respectively. The results revealed that protein extract of BFFW could induce Hela cells apoptosis.The apoptosis rate and the cytoclasis rate were by 28.075% and 18.3% in 72h respectively, which Hela cells were treated with 15.0μg/mL protein extract of BFFW. The results of Western Blotting revealed that included apoptosis pathway by BFFW was dependent on caspase activation.So, Fructus Trichosanth and Blaps femoralis Fischer- Waldheim had certain effect on Hela cell. They were the important research and development resources of new drug. It would provide theoretical and experimental basis for further development and clinical application of the above two kind of Chinese traditional medicine. |
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