| Choriocarcinoma is highly malignant trophoblastic tumor, whose rodent is stronger.Choriocarcinoma can transfer in early term in blood way, whose case fatality rate is very high. With the advancement of chemotherapy and the tumor's sensitivity to antineoplastic drug, greatly patients can be radical cured.Choriocarcinoma is one of the malignant tumor which can be cured by antineoplastic drug.However, multidrug resistance and recur is the main cause of the died patients of trophoblastic tumor chorioepithelioma. So, to control the multidrug resistance and tumor recur efficiently is very important for the tumor therapy . At present, the toxicity of reversal agent of drug resistance that have been explored limited its clinical application, and to restrain and block sing mechanisms of drug resistance can not eliminate MDR. The natural resources of Chinese medicinal materials is very abundant and, the traditional Chinese medicine can destroy the tumor cell by many target. At present, it is the focal point of tumor investigation to search new antitumor drug which is high performance . In our study, we discuss the three traditional Chinese medicine's effectiveness of reversion on drug resistance and their synergistic action with MTX. The study supply effective medicine and means for the therapy of drug fast chorioepithelioma. Objective: To determine whether the active constituent of three traditional Chinese medicine ( include shikonin tertrandrine and tanshinonl ) can resist the drug fast cell of chorioepithelioma, whether they have the effective of drug fast reversion. Methods: The experimental JAR/MTX cell were devided into four groups: ①Control groups: blank(simple cell) group, DMSO dissolvent group; ②chemotherapeutics group:cell+MTX; ③traditional Chinese medicine group:cell+monomer of traditional Chinese medicine; ④chemotherapeutics and traditional Chinese medicine group:cell+MTX+monomer of traditional Chinese medicine . We determine the toxicity of every medicine by MTT method and to obtain the IC50 of experiment medicine by it. Every medicine affect on the cell by concentration of C or 5C. After 12h 24h, we collect the cell and its clear supernatant liquid to observe the cell's appearance, to determine the HCG and to determine the expression of HSP27 and GST-π.. In addition, we determine the reversion multiple of every traditional Chinese medicine after they compound with MTX . Results: ⒈The toxic action of there traditional Chinese medicine and MTX : Every medicine's measured value of IC50 after on JAR/MTX were obtained.Shikonin's IC50 is about 2μg/ml, tanshinone's IC50 is about 4μg/ml, tetrandrine's IC50 is about 4μg/ml,MTX's IC50 is about 9.5μg/ml. It is thus evident that,above-mentioned medicine all can cause the JAR/MTX cell's death. But the MTX's IC50 is higher and obviously drugfast.ON the contrary, other three traditionary Chinese medicine's IC50 are lower , and are relatively sensitive . ⒉The morphology's change of JAR/MTX cell after every medicine affect the cell. ⑴MTX group: Normal JAR/MTX cell are polygon or irregularity,and inequality of size. Some cells have long ecptoma,JAR/MTX cell's adhesiveness is very high. We can see many caryon's mitotic figures. The change of DMSE dissolvent group hadn't obviouse distinguish with normal JAR/MTX cell group. After MTX affect on cells for 12h, the cells change are not obviously. The cells main change were transform to round,but remain the integrity on the whole, the proportion of nucleus/cytolymph are roughly normal.Most caryotheca were not disappear and lesser dead cell occurred, such as cellular swelling and breaking,cell envelope is disappearance. ⑵Traditional Chinese medicine group: Three traditional Chinese medicine such as Shikonin and soon all can obviously restrain the growth of JAR/MTX cell, and they can cause cellular necrosis. The effection of Shikonin and tertrandrine were prominent.After the effect of low dose and short short time of Shikonin and tertrandrine, the cells'sticking was breakdown, thecells shrinked and transformed to round; After the effect of medium dosage of Shikonin and tertrandrine, most cells amotic from coverslip and float in the culture fluid.We can saw a great quantity of breaking and dead cells. The cells'change of Shikonin group is caryon's swelling, coloretur's thinning, caryotheca's shrinkage and disappear and caryocinesis decrease. The cell's change of tetrandrine is cellular necrosis, such as nucleus condense,nucleus fragmentation; The cell death effect of tanshinone Ⅱa(C) is not obviously for 12h.With the lasting of time and raising of density, dead cells were increasing. Only when later period(24h), we can saw major dead cell. The decreasing of nucleus mitotic figure was not obviously. The main appearance were nucleus condense and nucleus fragmentation . ⒊The change of β-HCG level after the effect of every medicine The β-HCG of simple cell group had not significant deviation with DMSO dissolvant group(P>0.05). After three traditional Chinese medicine(C and 5C) affected on cell for 24h, the level of β-HCG decreased obviously compared with simple cell group and DMSO dissolvent group(P<0.05). So, all the threetraditional Chinese medicine can cut down the secretion of the JAR/MTX cells.Among the total, the effect of Shikonin is most signification, the next is tetrandrine, the last is tanshinone.It displayed the relationship of close-dependent; But the chemicals medicine MTX's effect (C) was not obviously, it's β-HCG had not obvious distinguish with normal cell groups(P>0.05). After the effect of MTX(5C), effectiveness can be saw(P<0.05) . ⒋The expression of HSP27 and GST-πof JAR/MTX cells after the effect of every medicine. All the normal JAR/MTX cells and DMSO dissolvent cell had higher expression of HSP27 and GST-πwas mainly in cytolymph and HSP27 was mainly in nucleus.All the three traditional Chinese medicine can decrease the expression of HSP27 and GST-π(P<0.05)[except the tanshinone Ⅱa of C had not evident distinction which compared with control group(P>0.05)]. The effect had relationship of close-dependent.Among these medicine, the effect of Shikonin is outstanding; But MTX only decreased the expression of GST-πon density 5C(P<0.05), and it can not decrease the expression of HSP27 whether on density C or on 5C(P>0.05). It is thus clear that all the rhree traditional Chinese medicine can decrease theexpression of HSP27 and GST-π.On the contrary, the effect of MTX is not obviously . ⒌The determination of three traditional Chinese medicine's sensitization effect. Three traditional Chineses medicine can derease the IC50 of MTX after they affect cells along with MTX. It indicated that every traditional Chinese medicine have noticeable efftct synergia with MTX. After affecting on cells along with Shikonin, IC50 of MTX is 1.8 μg/ml, Shikonin's reversal index is 5.3; After affecting on cells along with tetrandrine, IC50 of MTX is 2.3 μg/ml, tetrandrine's reversal index is 4.1; After affecting on cells along with tanshinone Ⅱa, IC50 of MTX is 3.6 μg/ml, tanshinone's reversal index is 2.6.Among these medicine,the synergism action of shikonin is extremely predominant, and tanshinone is weaker than other two traditional Chinese medicine . Conclusions: ⒈Active constituent of shikonin, tanshinone Ⅱa and tetrandrine all can restrain the growth of JAR/MTX cells, and can cause cellular necrosis.Among these medicine shikonin and tetrandrine's effects are most predominant.Cellular necrosisincrease with the raising of the medicine's density and prolonging of the effect time;Tanshinon Ⅱa's effect of celluar necrosis is obviously only on it's higher density and longer affecting time. So, the cellular necrosis have intimate relationship with medicine's density and effecting time . ⒉All the three traditional Chinese medicine can cut down the secretion of the JAR/MTX cells. Among the total, the effect of Shikonin is most signification , the next is tetrandrine, the last is tanshinone.It displayed the relationship of close-dependent; But the chemicals medicine MTX's effect (C) was not obviously, it's β-HCG had not obvious distinguish with normal cell groups. After the effect of MTX(5C), effectiveness can be saw . ⒊Three traditional Chinese medicine have obvious effect on JAR/MTX cells through many effect reversion pathway and different effect target, such as they have different down regulation on HSP27 and GST-π.. So, they can reverse drug fast efficiently . ⒋Three traditional Chineses medicine can derease the IC50 of MTX after they affect cells along with MTX . It indicated that every traditional Chinese medicine have noticeable efftct synergia with MTX . Among these medicine, the synergism...
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