The Research On Antitumor Activity Of Monomer Compounds From Cynanchum Chinense In Vitro

Objective To research the inhibition of three different monomer compoundsextracted from Cynanchum Chinense on tumor cells proliferation in vitro, andinvestigate the possible mechanisms.Method By CCK-8observation method, Using gastric mucosa GES-1cells asnormal contrast,With different concentrations of monomer compounds extractedfrom Cynanchum Chinense that are n-Octatriacontane(Monomer Compounds-Ⅱ,MC-Ⅱ),Kaempferol-7-O-α-L-rhamnose glycoside(Monomer Compounds-Ⅴ,MC-Ⅴ) and7-O-α-L-rhamnopyranosyl-kaempferol-3-O-α-L-rhamnose glycoside(Monomer Compounds-Ⅵ,MC-Ⅵ) respectively to act on human cervical cancerHela cells, mouse lymphoma cells YAC-1and human gastric cancer MGC-803cellline24h,48h,72h,96h,To observe the influence of three tumor cell inproliferation activity, And the hoechst33342/PI dye staining was carried out tocheck the apoptotic morphological changes for the tumor cells sensitive to allthree monomer compounds.Results①Hela cells were treated with MC-Ⅱ, and the values of OD betweendifferent dose groups varied from48h to72h. The proliferation of YAC-1cellscould be increased by MC-Ⅱ at24h,48h, and72h, while inhibited at96h; Helacells and YAC-1cells were treated with MC-Ⅴ and MC-Ⅵ at the concentration of50,100, and200μ g·mL-1respectively, as a result, the values of OD is lower inthe experimental groups than that in control groups(P<0.05),and is dose-depended. Similarly,24h,48h,72h, and96h after MGC-803cells were treated with differentdose of MC-Ⅴ, the values of OD is lower in the experimental groups than that incontrol groups(P<0.05), however, the inhibition effect of MC-Ⅵ on MGC-803cells could be observed only at24h after treated.②Hela, YAC-1and MGC-803cells were treated with different concentration of MC-Ⅴ at48h, the tumor cellswas observed under fluorescence microscope after Hoechst33342/PI staining,cells begin to shrink and nuclear pyknosis,Agglutination of chromatin Shows agrowing bright blue light and red light by the increasing concentration of MC-Ⅴ,the most obvious among them is gastric cancer cells MGC-803,72h after theKaryopyknosis of tumor cells was Formed obvious round bead and cell edgechange seriously, showing uneven coloration of the fluorescent chromogenicstructure.③YAC-1cells were treated with MC-Ⅵ at48h, the experimentalconcentration groups showed a growing blue light and red light,meanwhile,Karyopyknosis emit strong bright blue light,and is dose-dependent manner.Asmonomer drug induced by the extension of time for72h,the cell wall wasdestroyed seriously and uneven splitting cells increased, Nucleus with PI stainedgranular uneven shades of strong orange-red fluorescence and showed typical latecell apoptosis morphological structure.Conclusion①The experiments in vitro suggested that the growth of the tumorcells could be significantly inhibited by the monomer compounds extracted fromCynanchum Chinense compared to the growth of normal gastric mucosa GES-1cells, and the sensitivity of tumor cells is different.②The proliferation of Helacells, YAC-1cells and MGC-803cells could be obviously inhibited by flavonoidglycosides MC-Ⅴ isolated and purified from Cynanchum Chinense using Ethylacetate method, while only the YAC-1cells proliferation could be inhibited byflavonoid glycosides MC-Ⅵ isolated and purified from Cynanchum Chinense using n-butanol method.And they showed a time-and dose-dependent manner.③Noobvious anti-tumor activity was observed when the tumor cells were treated withthe alkane MC-Ⅱ isolated and purified from Cynanchum Chinense using Ethylacetate method.