| OBJECTIVE:Clinical Study objective: To investigate the effect of Jianpi Yishen Huatan Decoction on NLR,immune function and progression-free survival of patients with stage Ⅲ-Ⅳ lung squamous cell carcinoma treated by immunotherapy combined with chemotherapy or immunotherapy combined with arotinib.Experimental research objective: To study the effect of Jianpi Yishen Huatan Decoction on the proliferation and apoptosis of human lung squamous cell carcinoma SK-MES-1 cells,and observe whether Jianpi Yishen Huatan Decoction can down-regulate the expression of PD-L1 in SK-MES-1 cells,in order to seek evidence for the treatment of lung squamous cell carcinoma with Jianpi Yishen Huatan Decoction combined with immune checkpoint inhibitors.METHODS:Clinical study: Collect the medical records of 53 patients diagnosed as stage Ⅲ-Ⅳ lung squamous cell carcinoma in the Department of Oncology and Respiratory,Department of Thoracic Surgery and Outpatient Treatment of the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,record their progression-free survival time through regular followup,analyze the PFS of the two groups,use chi-square test to study the changes of NLR and lymphocyte count before and after treatment,andunderstand the changes of patients’ immune function,Finally,survival analysis was carried out by Kaplan-Meier curve and COX multifactor to study the influencing factors of patients’ PFS.P<0.05 represents a statistically significant difference.Experimental research: Prepare the medicated serum of Jianpi Yishen Huatan Decoction,culture the human lung squamous cell carcinoma SKMES-1 cells,count the cells and calculate the density,measure the cell viability with CCK8 method,use flow cytometry to detect the apoptosis rate of cells after treatment with Annexin V-FITC/PI method,and finally use Western blot method to detect the PD-L1 protein expression of cells after treatment with medicated serum.Results:Clinical study:1.Clinical efficacy: The median PFS of the treatment group(traditional Chinese medicine + immunotherapy combined with chemotherapy or immunotherapy combined with arotinib)was 8.40 ± 0.35 months,and the median PFS of the control group(immunotherapy combined with chemotherapy or immunotherapy combined with arotinib)was 7.20 ± 0.42 months.The median PFS of the two groups was tested by Log-rank test,and the difference was statistically significant(P=0.006<0.01).In subgroup analysis,it was found that there were significant differences in PFS between the two groups of patients with different treatment schemes,regardless of whether they were treated with traditional Chinese medicine.The PFS of the immunotherapy combined with arotinib group was better than that of the immunotherapy combined with chemotherapy group(P=0.000,P=0.001 <0.01).2.Influencing factors of PFS: Patients with low or undifferentiated pathological differentiation have a worse prognosis than those with mediumand high differentiation(median PFS 5.0±0.39 months vs 8.30±0.25 months,CI 95% 12.321-204.063,P=0.000<0.05).The degree of pathological differentiation is an independent adverse prognostic factor of PFS.3.Comparison of NLR before and after treatment: There was no significant difference in NLR level between the treatment group and the control group before receiving treatment(3.37±0.51 vs 3.48±0.52,P=0.446>0.05),and the NLR level in the treatment group was significantly lower than that in the control group after treatment(3.09±0.41 vs 3.51±0.42),with a statistically significant difference(P =0.001<0.01).4.Effect on immune function: There was no significant difference in CD3+,CD4+,CD8+ and CD4+/CD8+ between the treatment group and the control group before receiving treatment(P>0.05 for each comparison group).After treatment,the ratio of CD3+,CD4+ and CD4+/CD8+ in the treatment group were significantly higher than those in the control group(P<0.01 for each comparison group),while CD8+ was significantly lower than that in the control group(P<0.01).Experimental research:CCK8 assay was used to detect the proliferation of SK-MES-1 cells treated with low,medium and high doses of Jianpi Yishen Huatan Decoction medicated serum for 24 h.Oneway-ANOVA univariate statistics showed that Jianpi Yishen Huatan Decoction could significantly inhibit the proliferation of SK-MES-1 cells,and the effect was directly proportional to the drug concentration;flow cytometry showed that after 24 h of intervention with different concentrations of Jianpi Yishen Huatan Decoction,using Oneway-ANOVA univariate statistics,different concentrations of Jianpi Yishen Huatan Decoctioncould induce apoptosis,of which the apoptosis rate of medium and high dose medicated serum groups increased significantly;Western blot assay suggested that high dose of Jianpi Yishen Huatan Decoction medicated serum intervened cells,which could decrease the PD-L1 protein expression of cells,and when combined with anti-PD-1 antibody drugs,the PD-L1 protein expression of cells was significantly lower than the control group.Conclusion:1.This study shows that Jianpi Yishen Huatan Decoction can prolong the PFS of patients with advanced squamous cell carcinoma of the lung;The PFS of the immunotherapy combined with arotinib group is better than that of the immunotherapy combined with chemotherapy group,and the combined treatment with Chinese medicine may have better curative effect.2.Jianpi Yishen Huatan Decoction can significantly reduce the NLR value of LCSS patients’ peripheral blood,and can increase the proportion of CD3+,CD4+ and CD8+,improve the cellular immune function,enhance the immune response ability of patients to tumor cells,increase the number of lymphocytes,increase the CD4+/CD8+ value,and reverse the immunosuppressive state of the body.3.The serum containing Jianpi Yishen Huatan Decoction has a significant inhibitory effect on the proliferation of lung squamous cell carcinoma SK-MES-1 cells and can effectively induce apoptosis of lung squamous cell carcinoma SK-MES-1 cells,and the utility of both of them is positively correlated with the drug concentration.4.Jianpi Yishen Huatan Decoction reduced PD-L1 protein expression levels in lung squamous cell carcinoma SK-MES-1 cells,and PD-L1 protein decreased more significantly when combined with tislelizumab monotherapy. |
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