| Background:Lung cancer is now the most often encountered malignant tumor in the world. Eighty-percent of lung cancer is non-small-cell lung cancer that is number one in both its occurrence and death rate within urban populations. Two thirds of these cases occur in senior patients as late stage lung cancer. Cisplatin is officially recognized as the number one drug for treating lung cancer, but due to cisplatin's cellular toxicity, clinical usage of the drug results in numerous severe side-effects. These severe side-effects often make the cisplatin treatment impractical for senior patients. "Invigorating Spleen & Phlegm Elimination" method supports the healthy energy as its main approach, and has seen widespread usage in clinics. Investigation into its effects in decreasing cisplatin's toxicity and increasing cisplatin's function has important clinical implications which may expand cisplatin's possible clinical applications.Objective:Using TCM "Invigorating Spleen & Phlegm Elimination" herbs to treat NSCLC tumor both in vivo and in-vitro, in order to appraise its efficacy in NSCLC cancer treatment. To observe the survival rate and the tumor Growth Inhibition rate of TCM "Invigorating Spleen & Phlegm Elimination" only treatment, the cisplatin only treatment, and the combination treatment with both cisplatin and TCM. The comparison of these data will disclose the anti-cancer effects of TCM "Invigorating Spleen & Phlegm Elimination" and its effectiveness to reduce the cisplatin toxicity and increase survival rate of the cisplatin's treatment. The result will be only an initial exploration into the effects and mechanism of the combination treatment, which may serve as reference for creating reasonable combination treatment plans involving both cisplatin and TCM.Methodology:In-vivo cell experiment:The MTT method is used to identify the most appropriate LL/2 cell count for the serial in-vivo experiment. Then, the effects of different concentrations of "Invigorating Spleen & Phlegm Elimination" and cisplatin on LL/2 cell vitality are measured.In-vitro animal experiment:First, the optimal dosage of cisplatin has been confirmed experimentally, which has twelve mice C57BL/6J implanted with LL/2 tumor cell as subjects, divided into Control group,5 mg,6 mg, and 7 mg groups with three members each. After drug treatments, the mice are examined and measured for tumor size and body weight every 3-4 days and terminated on the 15th day. The latter combination study of cisplatin with TCM has twenty-four mice C57BL/6J implanted with LL/2 tumor cell as subjects treated with the optimal dosage of cisplatin, divided into the Control group, TCM p. o. group, cisplatin i.p. group, TCM p. o.+cisplatin i. p. group, with six members each. After drug treatment the mice are examined and measured for tumor size and body weight every day and terminated on the 21st day. The hearts, livers, lungs, spleens, kidneys, and cancerous tissues are removed, weighed, sectioned and dyed with HE in order to observe changes.Results:In-vivo cell experiment:The optimal cell count of LL/2 cells for MTT experimentation is 3000 per well (on 96 well plates). Less than 200 ug/ml of TCM "Invigorating Spleen & Phlegm Elimination" has no effect on LL/2 cell vitality. Greater than 10uM of cisplatin clearly inhibits LL/2 cell regeneration, accelerating tumor cell apoptosis.In-vitro animal experiment:Cisplatin's optimal dosage is 5mg/kg. The results of TCM and cisplatin combination treatment are as follows.1. General condition:Nodules approximately the size of half a rice grain are palpated at the implantation site on the lateral aspects of right hind leg 5 days after LL/2 tumor cell implantation. Subsequently, these nodules gradually increased in size. The tumor growth tendencies of Cisplatin i.p. group and those groups receiving TCM treatment are both less than that of the control group. Cisplatin i. p. group and those groups that received abdominal cisplatin injections all show significantly less growth than the control group. In comparison to the control group, the mice in all other groups show lower tumor growth rate and growth margin.2. Tumor suppression effect:Tumor volume:Control group (3757.64±2222.95)> TCM po group (2622.18±1567.18)> TCM po±Cisplatin ip group (2170.40±871.60)> Cisplatin ip group (1311.81±655.55).Tumor weight:Control group (3.4392±1.4857)> TCM po group (2.4205±1.3809)> TCM po+Cisplatin ip group (2.3716±0.9884)> Cisplatin ip group (1.4295±1.1208).Tumor suppression rate:Cisplatin ip group (58.43%)> TCM po+Cisplatin ip group (31.04%)> TCM po group (29.62%)3. Survival rate:26 day survival rate:TCM po+Cisplatin ip group (83.30%) > TCM po group (66.67%)> Cisplatin ip group (50.00%)= Control group (50.00%)4. Histology section results: Heart, liver, spleen, lung, kidney, and tumor sections of each group are observed under the microscope. The changes in spleen and tumors are readily apparent. The changes in other organs are not apparent.a) Spleen section examination:T cell≥B cell rate in spleen:TCM po group (75%)> TCM po+Cisplatin ip group (60%)> Control group (33.33%)> Cisplatin ip group (0%)b) Tumor necrosis rate:Cisplatin ip group (100%)> TCM po+Cisplatin ip group (80%)> TCM po group (25%)> Control group (0%) Conclusion:TCM "Invigorating Spleen & Phlegm Elimination" has suppressive effects on LL/2 lung cancer, and can increase survival rate. The mechanism may involve promoting T-cell regeneration, and increasing the body's immune capabilities. Although the combination treatment with TCM "Invigorating Spleen & Phlegm Elimination" and Cisplatin did not improve Cisplatin's tumor-suppression effects, the combination treatment clearly increased survival rate beyond that of using each drug alone. TCM alleviating Cisplatin's immuno-suppressive effects is a possible mechanism.
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