| Objective: Cervical cancer is one of the most common gynecological tumors, about 200 thousands women die of cer -vical cancer all over the world every year. Cervical cancer seriously threatens women's health and the quality of their lives. In recent years, on the basis of traditional therapy methods,the study of immune therapy is increasingly becoming a hot issue for scholars all over the world. Tumor can escape from immunological surveillance through immunosuppression caused by immunosuppressive mole cules secreted from tumor cells, and this is the immportant mechanism of the occurrence and development of tumors. The supernatant immunosuppression from cultured Hela cells formed by secreted immunosuppressive molecules in which TGF-β1, IL-10 and PGE2 were reported more. Therefore, studying the down-regulative effects on tumor immunosuppressionand immunosuppressive molecules secretion and seeking for the effective drugs that can reverse this kind of immunosuppression are important parts of discovering anti-tumor drugs, ascertaining their immunological mechanisms and guiding their proper clinical application. The study of anti-tumor traditional Chinese medicines had dominant position in the area of study on anti-tumor drugs. The study used Hela cells that has effects of immunosuppression from supernatant, followed by treated with three anti-tumor traditional Chinese medicines respectively, to observe the effects of re-cultured supernatant on immunosuppression and the concentration of immunosuppressive molecules in the supernatant. It is expec -ted to provide experimental basement and especially charac -teristic anti-tumor traditional Chinese medicines that have our own property right and guide the proper application for clinical uses.Methods: the suitable inoculated concentration and cultured time of Hela cells were detected by MTT. Re-cultured supernatant of Hela cells after treated with Ligustrazine Hydrocholoride (LHC), Matrine N-Oxide (MOX) and Artemether (ART) respectively for 48 hours(the top of the non-poisonous concentration has been determined by MTT), the supernatants were called LHC-S, MOX-S and ART-S. Followed by being washed completely to remove the traditional Chinese medicines, and re-cultured twice with fresh medium , the supernatants we collected were called LHC-S1, MOX-S1, ART-S1 and LHC-S2, MOX-S2, ART-S2. The corresponding supernatants of Hela cells treated without any traditional Chinese medicines as control ,they were called control-S, control-S1 and control-S2. The concentrations of three immunosuppressive molecules (including TGF-β1, IL-10 and PGE2 ) in different supernatant were measured by quantitative ELISA (TGF-β1 and IL-10 ) and ultraviolet spectrophotometry A278(PGE2). The statistics analysis were used between the immunosuppression and concentrations of immunosuppressive molecules from re-cultured supernatants of Hela cells after treated with traditional Chinese medicines, in order to suspect their relationship, to ascertain the corresponding target molecules of each traditional Chinese medicine that can down-regulate the immunosuppression from Hela cells.Results: 1 The inoculated concentration were deter -mined by MTT: 2×105/mL. 2 The top of the non-poisonous concentration of the three medicines were: LHC7.80mg/mL,MOX 39.06mg/mL, ART 3.13mg/mL. 3 For supernatants of Hela treated without any traditional Chinese medicines, all of the 3 immunosuppressive molecules were secreted and were found steadily, the concentration of TGF-β1 was highest, which was about (989.226±34.836)ng/mL, the concentration of IL-10 was( 3.768±0.145) pg/mL , A278 of PGE2 was (0.262±0.037 ); for the Hela cells treated with LHC, the concentration of TGF-β1 , IL-10 and PGE2 in LHC-S decrea -sed greatly( 18.791±2.368, P<0.001; 2.982±0.117, P<0.05; 0.148±0.037, P<0.05 ). Compared with control-S in which the decreased degree of TGF-β1 was most, and its concentr -ation was only 1.90% of control-S,the concentration of IL-10 was 87.90% of control-S, the A278 of PGE2 was 56.49 % of control-S;compared with LHC-S, the concentration of TGF-β1 still decreased greatly(2.398±0.458, P<0.01), was only 0.25% of control -S1, IL-10 had no changes (2.871±0.018,P>0.05),PGE2 still decreased greatly(0.091±0.007, P <0.01), was 37.73% of control-S1; compared with LHS1, the concentration in LHC-S2 of TGF-β1 continue to decrease(0.595±0.031,P <0.001), was only 0.06% of control-S2, the concentration of IL-10 in LHC-S2 increased highly(3.232±0.116,P<0.05),but less than the concerntration of control-S2, A278 of PGE2 begin to increase greatly (0.144±0.042, P<0.05) . For Hela cells treated with MOX, the concentrations of TGF-β1 in MOX-S decreased greatly (2.561±0.78 9,P<0.001),only was 0.26% of control-S ,IL-10 and PGE2 A278 had no changes(3.304±0.085, P>0.05; 0.215±0.057, P>0.05); compared with MOX-S, the concentration of TGF -β1 in MOX-S1 continue to decrease greatly(0.305±0.035, P<0.01), only was 0.03% of control-S1, IL-10 and PGE2 had no changes(3.492±1.008, P>0.05; 0.186±0.045, P>0.05); compared with MOX-S1, the concentration of TGF-β1 in MOX-S2 increased highly (1.382±0.389, P<0.001), was 0.14 % of control-S2, IL-10 and PGE2 had no changes(3.047±0.239, P>0.05; 0.151±0.035, P>0.05). For Hela cells treated with ART, the concentrations of TGF-β1 decreased greatly in ART-S(2.859±0.520, P<0.001), only was 0.29% of control-S, the concentrations of IL-10 decreased greatly(3.010±0.217, P<0.05), too, was 88.76% of control-S, PGE2 decreased greatly(0.149±0.013, P<0.05) ,was 58.43% of control-S; com -pared with ART-S, the concentration of TGF-β1 in ART-S1 continue to decrease(0.009±0.001, P<0.001), only was 0.000 08% of control-S1,the concerntration of IL-10 decrease -d (3.121±0.137, P<0.05), was 90.99% of control-S1, PGE2 decreased greatly (0.138±0.032, P<0.05), was 52.67% of control-S1; compared with ART-S1, the concentration of TGF-β1 in ART-S2 increased highly(0.576±0.032, P<0.001 ), was 0.06% of control-S2, the concentration of IL-10 and PGE2 had no changes(3.207±0.082, P>0.05; 0.186±0.013, P>0.05).Conclusions:1 Hela cells secreted the 3 immunosuppr -essive molecules detected steadily. 2 The three traditional Chinese medicines LHC,MOX and ART down-regulated the concentrations of TGF-β1 secreted by Hela cells ,especially ART; LHC and ART can down -regulated the secretion of IL-10 and PGE2 ,especially LHC. 3 TGF-β1 may be the dominant immunosuppressive mole -cules secreted by Hela cells. 4 Down-regulate the secretion of immunosuppressive moles may be the new anti-tumor mechanism of LHC,MOX and ART.
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