| Cervical cancer is the most common gynecological malignant tumor and the second largest cancer causing death in womenMeanwhile cervical cancer is still lack of fully effective treatment at present.The traditional radiotherapy and chemotherapy cannot reduce mortality,the inevitable toxic and side-effects reducing the patient's quality of life greatly.Now we believe it is because of immuno-suppressive factors existing in the tumor micro environment.Tumor micro environment in tumor which cells can secrete some immune inhibitory factors such as TGF-beta 1,IL-10.So how to reduce or eliminate the immunosuppressive factor in tumor micro-environment and enhance the treatment effect,this important problem is an urgent need to solve in tumor therapy.Lactobacillus is a group of normal advantage bacterium in woman's vagina,which plays a vital role in maintaining vaginal normal ecological balance.Several studies have shown that cervical cancer is associated with vaginal micro-ecological balance disorders.The lactobacillus decrease,which causes vaginal Gardner or mixed anaerobic bacteria to speed up its proliferation and produce harmful substances,together with other carcinogenic factors(such as the human papilloma virus)jointly affected the cervix,accelerated the development of cervical cancer.Whether we can reduce or eliminate the immunosuppressive factor in tumor microenvironment and enhance the treatment effect with combined lactobacilli with cisplatin for treatment of cerical cancer,we will be provided a new prospective by this study.Methods: 1 Experiment in vitro 1.1 Develop cervical cancer U14 cells and lactobacilli.1.2 By using cisplatin,lactobacillus,cisplatin + lactobacillus,we could stimulate U14 cervical cancer cells,controled of PBS.24 h,48 h,72 h cells collection,and observed U14 cells TGF-beta 1,IL-10 mrna expression.2 In vivo testsForty 4~6 weeks of female C57BL/6 mice,were randomly divided into 4 groups,each group had 10 mice.The 2x10~6/0.2ml U14 cells were then subcutaneously injected into the C57BL/6 mice.Tumor-inhibiting experiment was conducted after 7 days after tumor-burdened.In lactic acid bacteria group,each mouse was only given intraperitoneally with lactobacilli 2.25*10~6/0.2 ml arround diffirent parts of tumor;in cisplatin group,0.2ml cisplatin was given intraperitoneally to each mouse;in cisplatin and lactic acid bacteria group,each mouse was given intraperitoneally with 0.2 ml cisplatin and with lactobacilli 2.25*10~6/0.2ml arround diffirent parts of tumor,the control group injected the same amount of PBS,once per day for 14 days.Checked mice weight and the size of the tumor everyday.After a day's withdrawal,executed mice.Weighed tumors and used vernier calipers to measured tumors length diameter(L)and width diameter(W),and to calculated the tumors volume according to the formula V=L.W2/2.Cut the tumors,depicted the tumors growth curves,observed its section,internal texture,calculated the tumors.Divided tumor tissue by using 10% paraformaldehyde fixed liquid to fixation to do immune-histo-chemical detection the above factors.Results:1 In vitro experiments: Forty-eight hours after the corresponding stimulation in cervical cancer U14 cells,on the mRNA levels,cisplatin group,combined treatment group were significantly less than Lactobacillus group and control group in expression quantity of TGF-beta 1,IL-10 respectively,differences were statistically significant;cisplatin group were significantly less than lactobacillus group and control group in expression quantity of TGFbeta 1,IL-10 respectively(P<0.05),differences were statistically significant;combined treatment group were significantly less than lactobacillus group and control group in expression quantity of TGF-beta 1,IL-10 respectively(P<0.05),differences were statistically significant;cisplatin group were significantly higher than combined treatment group in expression quantity of TGF-beta 1,IL-10 respectively(P<0.05);there were no statistical difference been found between the comparison between control group and lactobacillus group(P>0.05).2 In vivo experiments(The growing status of each tumor-burdened mouse): First 10 days after tumor-burdened,every group of mice grew in good condition,but 15 days later,the differeces began to appare,the weight of control group mice began to decline,and these mice became thinner and less active;7 days later from been given intraperitoneally,cisplatin group mice appeared to have no appetite;14 days after,they became bradycardia and their spirits were drooping;while the mice of combined treatment group and the mice of lactobacillus group were in good condition,their weight did not significantly decline.3 In vivo experiments(The growing status of the tumor in mice and calculation of the tumor inhibitory rate):First 9 days after inoculation of tumor cells,each group was all good;10 to 14 days,the control group mice tumor began to grow faster than other three groups,among which the slowest was combined treatment group;15 to 24 days,the control group's tumor growth in mice were significantly faster than other three groups;and cisplatin group and combined treatment group mice tumor became smaller,the slowest tumor growth is combined treatment group.The weight of control group,the lactic acid bacteria group,cisplatin group and combined treatment group tumor,were(3.158±0.004)g,(2.901±0.005)g,(2.070±0.004)g and(1.576±0.003)g respectively,The control group tumor weight is bigger than others.The multiple comparisons had been carried out in tumor weight among the different groups,the data were statistically different;calculation based on the inhibitory rate formula,lactic acid bacteria group,the combined group,cisplatin group inhibitory rate were 8%,50%,34%,the highest inhibitory rate was combined treatment group,the least is the control group.4 Immunohistochemical results in vivo experiments: in the control group,the lactic acid bacteria group,combined treatment group and cisplatin group,each of TGF-beta 1 average optical density value was(0.466±0.003,(0.438±0.002),(0.122±0.005)and(0.144±0.001);Each of IL-10 average optical density value was(0.480±0.005),(0.472±0.003),(0.128±0.002)and(0.152±0.006);the multiple comparisons had been carried out,cisplatin group,combined treatment group were significantly less than lactobacillus group and control group in expression quantity of TGF-beta 1,IL-10 respectively,differences were statistically significant;cisplatin group were significantly less than lactobacillus group and control group in expression quantity of TGFbeta 1,IL-10 respectively(P<0.05),differences were statistically significant;combined treatment group were significantly less than lactobacillus group and control group in expression quantity of TGF-beta 1,IL-10 respectively(P<0.05),differences were statistically significant;cisplatin group were significantly higher than combined treatment group in expression quantity of TGF-beta 1,IL-10 respectively(P<0.05);there were no statistical difference been found between the comparison between control group and lactobacillus group(P>0.05).Conclusions: The status of combined group mice were better than the control group,and were smaller than the control group in the tumors volume obviously.Cisplatin combined with lactobacilli could be tested in a tumor-burdened mice,they can cut immunosuppressive factor in tumor tissue expression of TGF-beta 1 and IL-10,relieve the immune suppression in the tumor micro environment.Relative to use cisplatin alone,combined lactobacilli with cisplatin for treatment of cerical cancer would be higher the treatment effect and less toxic and side effects.So combined treatment would be a great research value of clinical application. |
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