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Table of Content

    08 May 2020, Volume 47 Issue 5 Previous Issue    Next Issue
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    Original Articles
    Overexpression of miR-125 inhibits proliferation and invasion of nasopharyngeal carcinoma cells by down-regulating FGF-2
    Yang Chao, Wang Jianbo, Zhang Juhong, Wang Jialong
    2020, 47 (5):  257-263.  doi: 10.3760/cma.j.cn371439-20191114-00015
    Abstract ( 493 )   HTML ( 27 )   PDF (1873KB) ( 234 )   Save

    Objective To explore the expression of miR-125 in nasopharyngeal carcinoma tissues and the possible regulatory mechanism of biological characteristics of tumor cells. Methods Thirty cases of carcinoma and paracancerous tissues from patients with nasopharyngeal carcinoma admitted to Southern Hospital of Sixth People's Hospital Affiliated to Shanghai Jiaotong University from June 2018 to June 2019 were collected. The expressions of miR-125 and fibroblast growth factor 2 (FGF-2) mRNA were detected by fluorescent quantitative PCR. Nasopharyngeal carcinoma CNE-2 cells were transfected by miR-125 mimic (miR-125 mimic group), and negative control group was set (NC group). Transwell chamber assay was used to determine cell invasion ability, scratch healing assay was used to determine cell migration ability, WST-1 assay was used to assess cell viability, flow cytometry and electron microscopy were used respectively to detect apoptosis and autophagy, dual luciferase reporter assay was used to analyze the target of miR-125, and Western blotting was used to detect the expressions of related proteins. Results The relative expression of miR-125 mRNA in nasopharyngeal carcinoma tissues was 0.692±0.316, which was significantly lower than 1.501±0.748 in the adjacent tissues (t=5.242, P<0.001). The relative expression of FGF-2 mRNA in nasopharyngeal carcinoma tissues was 1.317±0.552, which was significantly higher than 0.783±0.241 in the adjacent tissues (t=7.360, P<0.001). The miR-125 mRNA expression of CNE-2 cells in the miR-125 mimic group was 4.091±0.145, which was significantly higher than 0.993±0.137 in the NC group (t=85.062, P<0.001). The proliferative activities of CNE-2 cells in the miR-125 mimic group at 48 and 96 h after transfection were significantly lower than those in the NC group (0.891±0.214 vs. 1.295±0.245, t=6.802, P<0.001; 0.934±0.208 vs. 1.488±0.269, t=8.924, P<0.001). The number of transmembrane cells and cell migration rate of the miR-125 mimic group were 36 000±3 820 and (39.4±6.5)%, which were significantly lower than 74 000±7 500 and (102.7±10.6)% of the NC group (t=24.728, P<0.001; t=27.883, P<0.001). The apoptosis rate of CNE-2 cells in the miR-125 mimic group was (22.5±1.4)%, which was significantly higher than that in the NC group (1.4±0.5)% (t=77.740, P<0.001). The relative expression of the apoptotic protein Bax in the miR-125 mimic group was 0.983±0.158, which was significantly higher than that in the NC group (0.418±0.122; t=15.503, P<0.001), and the relative expression of Bcl-2 was 0.688±0.174, which was significantly lower than that of the NC group (1.013±0.109; t=8.670, P<0.001). Autophagy was observed in miR-125 overexpressing CNE-2 cells by electron microscopy, and the relative expression ratio of autophagy protein LC3Ⅱ/LC3Ⅰ in the miR-125 mimic group was 2.517±0.209, which was significantly higher than 1.238±0.135 in the NC group (t=28.156, P<0.001). The expression of FGF-2 protein in the miR-125 mimic group was 0.504±0.118, which was significantly lower than 1.228±0.134 in the NC group (t=22.210, P<0.001). The double luciferase report confirmed FGF-2 as the target gene of miR-125. At 12, 24, 48 and 96 h after the transfection, the cell proliferative activities of CNE-2 cells co-transfected by FGF-2 gene plasmid and miR-125 mimic were significantly higher than those of CNE-2 cells transfected by miR-125 mimic (all P<0.05), and the apoptosis rate was significantly lower than that of CNE-2 cells transfected by miR-125 mimic [(6.2±1.5)% vs. (17.6±2.4)%, t=22.062, P<0.001]. Conclusion The expression of miR-125 is down-regulated in nasopharyngeal carcinoma tissues. Overexpression of miR-125 may inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma CNE-2 cells and promote the apoptosis and autophagy of CNE-2 cells by down-regulating FGF-2 expression.

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    Expressions of LSD1 and PDPN in tongue squamous cell carcinoma and their effects on prognosis
    Wang Kunlong, Zhang Yang, Su Weipeng, Liu Pan, Zhao Huarong
    2020, 47 (5):  264-271.  doi: 10.3760/cma.j.cn371439-20200221-00016
    Abstract ( 540 )   HTML ( 24 )   PDF (1509KB) ( 324 )   Save

    Objective To investigate the expressions of lysine specific demethylase 1 (LSD1) and podoplanin (PDPN) in tongue squamous cell carcinoma and the correlation between LSD1 or PDPN and clinicopathological characteristics or prognosis. Methods A total of 67 cases of tongue squamous cell carcinoma and corresponding paracancerous normal tissues in the First Affiliated Hospital of Xinjiang Medical University from January 2011 to January 2016 were selected. The expressions of LSD1 and PDPN in cancer and paracancerous tissues were detected by immunohistochemical method, and the patients were followed up for a long time to analyze the correlation between the expression of LSD1 or PDPN and clinicopathological characteristics or prognosis. Results The expressions of LSD1 and PDPN in tongue squamous cell carcinoma tissues were higher than those in paracancerous tissues, and the differences were statistically significant (Z=6.089, P<0.001; Z=5.781, P<0.001). The expression intensities of LSD1 and PDPN were significantly different in patients with different clinical stage (χ 2=11.487, P=0.001; χ 2=8.111, P=0.004), lymph node metastasis (χ 2=4.772, P=0.029; χ 2=6.206, P=0.013) and tumor size (χ 2=5.985, P=0.014; χ 2=4.247, P=0.039). The expression intensity of LSD1 was also significantly different in patients with different degrees of differentiation (χ 2=6.660, P=0.010). In univariate analysis, LSD1 expression intensity was negatively correlated with progression-free survival (PFS) and overall survival (OS) (χ 2=18.930, P<0.001; χ 2=16.257, P<0.001), PDPN expression intensity was negatively correlated with PFS and OS (χ 2=31.720, P<0.001; χ 2=18.390, P<0.001), and tumor size was negatively correlated with PFS and OS (χ 2=5.326, P=0.021; χ 2=8.843, P=0.003). Postoperative radiotherapy and clinical stage were positively and negatively correlated with OS respectively (χ 2=4.197, P=0.040; χ 2=6.355, P=0.012). In multivariate analysis, LSD1 was an independent risk factor for PFS and OS (HR=5.743, 95%CI: 1.012-32.579, P=0.048; HR=17.759, 95%CI: 2.303-136.916, P=0.006), PDPN was an independent risk factor for PFS (HR=4.380, 95%CI: 1.258-15.254, P=0.020), postoperative radiotherapy was a protective factor for PFS and OS (HR=0.374, 95%CI: 0.157-0.895, P=0.027; HR=0.218, 95%CI: 0.091-0.521, P=0.001), and clinical stage was an independent risk factor for OS (HR=2.637, 95%CI: 1.107-6.280, P=0.029). In tongue squamous cell carcinoma tissues, the expression of LSD1 was positively correlated with that of PDPN (rs=0.655, P<0.001). Conclusion The expressions of LSD1 and PDPN in tongue squamous cell carcinoma are higher than those in adjacent tissues. LSD1 is an independent risk factor for PFS and OS, PDPN is an independent risk factor for PFS, clinical stage is an independent risk factor for OS, and postoperative radiotherapy is a protective factor for PFS and OS. There is a positive correlation between the expressions of LSD1 and PDPN in tongue squamous cell carcinoma, and they can both be used as independent predictors of prognosis in patients with tongue squamous cell carcinoma.

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    Efficacy of β-elemene combined with gefitinib in the treatment of advanced lung adenocarcinoma patients with slow progression after first-line gefitinib treatment
    Song Chunqing, Zhang Ying, Zhang Zhiguo, Guo Hongwei, Lu Hanhong, Han Lei
    2020, 47 (5):  272-277.  doi: 10.3760/cma.j.cn371439-20191112-00017
    Abstract ( 586 )   HTML ( 15 )   PDF (809KB) ( 250 )   Save

    Objective To investigate the efficacy of β-elemene combined with gefitinib in the treatment of advanced lung adenocarcinoma patients with slow progression after first-line gefitinib treatment and its effects on quality of life and immune function. Methods A prospective cohort study design was used to enroll patients with advanced lung adenocarcinoma who met the inclusion criteria from June 2017 to December 2018 in our hospital. They were divided into experimental group and control group by random number table method. The experimental group was given elemene combined with gefitinib, and the control group was only given gefitinib. The clinical efficacy, quality of life and immune function of the two groups were compared after 2 cycles of treatment. The PFS2 (time from slow progression of disease to rapid progression of disease) was followed up. Results A total of 61 patients were included. There were 30 cases in the experimental group and 31 cases in the control group. The disease control rates of the experimental group and the control group were 83.3% (25/30) and 58.1% (18/31) respectively, and the difference was statistically significant (χ 2=4.680, P=0.031). The short-term efficacy of the experimental group was better than that of the control group, and the difference was statistically significant (Z=-2.623, P=0.009). The median PFS2 of the experimental group was 4.20 months (95%CI: 3.94-4.46), and the median PFS2 of the control group was 4.00 months (95%CI: 2.94-5.07), with a statistically significant difference (χ 2=4.948, P=0.026). Quality of life was compared between the two groups: in terms of physical function, emotional function and overall quality of life, score differences before and after treatment of the experimental group were higher than those of the control group, with statistically significant differences [6.67(-6.66, 20.00) vs. 0(-6.66, 6.66), Z=-2.429, P=0.015; 29.17(2.08, 56.26) vs. 12.49(-14.59, 39.57), Z=-2.263, P=0.024; 16.67(-33.33, 56.67) vs. 8.34(-18.74, 35.42), Z=-2.249, P=0.025]. In terms of immune function, CD4 +T cells level in the experimental group increased after treatment compared with that before treatment (44.27%±6.78% vs. 41.17%±3.73%, t=-2.426, P=0.022), and CD8 +T cells level decreased compared with that before treatment (21.47%±3.18% vs. 23.50%±2.37%, t=2.532, P=0.017). After treatment, the level of CD4 +T cells in the experimental group was significantly higher than that in the control group (44.27%±6.78% vs. 39.63%±5.80%, t=2.725, P=0.011). Conclusion β-elemene combined with gefitinib has a certain effect in the treatment of advanced lung adenocarcinoma patients with slow progression after first-line gefitinib treatment, and the quality of life and immune function are improved. It is worthy of further clinical research.

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    Value of nomogram incorporated preoperative tumor volume on predicting the prognosis of thoracic esophageal squamous cell carcinoma patients
    Guo Xinwei, Ji Shengjun, Ji Lei, Zhang Han, Zhou Shaobing, Liu Yangchen
    2020, 47 (5):  278-283.  doi: 10.3760/cma.j.cn371439-20190930-00018
    Abstract ( 579 )   HTML ( 17 )   PDF (1376KB) ( 274 )   Save

    Objective To explore the influence of clinicopathological factors besides TNM stage, including preoperative tumor volume, length and maximum diameter, on survival prognosis of patients with thoracic esophageal squamous cell carcinoma (ESCC), and to evaluate the predictive survival rate of clinicopathological variables with statistical significance by nomogram. Methods A total of 296 patients with ESCC treated by radical resection at the Department of Thoracic Surgery of Affiliated Taixing People's Hospital of Yangzhou University from 2011 to 2014 were retrospectively analyzed. These patients were grouped for further analysis according to the optimal threshold of preoperative tumor volume, length and maximum diameter. Kaplan-Meier method was used to calculate survival rate and survival comparison was performed by log-rank test. The univariate and multivariate Cox models were used to analyze the relationships between clinical variables and survival prognosis. Finally, nomogram model was established by integrating statistically significant clinicopathological parameters, and the predictive value of this model was further verified by calibration curve, concordance index (C-index) and decision curve. Results The optimal thresholds of preoperative tumor volume were 32 cm 3 and 72 cm 3 by X-tile analysis, and among the patients whose tumor volume was <32 cm 3 (n=94), the 1-, 3- and 5-year survival rates were 100%, 84.0% and 68.1%; in the 32-72 cm 3 group (n=118), the 1-, 3- and 5-year survival rates were 98.3%, 42.4% and 24.6%; in the >72 cm 3 group (n=84), the 1-, 3- and 5-year survival rates were 94.1%, 25.0 and 7.1% (χ 2=86.639, P<0.001). The optimal cutoff values of tumor length were 3.0 cm and 5.0 cm, and among the patients with tumor length <3.0 cm (n=62), the 1-, 3-, and 5-year survival rates were 99.5%, 87.1% and 69.4%; in the 3.0-5.0 cm group (n=146), the 1-, 3-, and 5-year survival rates were 98.6%, 47.9% and 30.1%; in the >5.0 cm group (n=88), the 1-, 3-, and 5-year survival rates were 94.3%, 29.6%, 13.6%, respectively (χ 2=53.607, P<0.001). The thresholds of tumor maximum diameter were 2.5 cm and 3.5 cm, and among these, the 1-, 3- and 5-year survival rates were 99.5%, 84.3% and 74.5% in the maximum diameter <2.5 cm group (n=51); 98.3%, 57.0% and 36.4% in the 2.5-3.5 cm group (n=121); and 96.0%, 29.0% and 13.7% in the maximum diameter >3.5 cm group (n=124, χ 2=62.109, P<0.001). In univariate analysis, the following factors were significantly associated with overall survival (OS): tumor location, differentiation grade, T stage, N stage, TNM stage, adjuvant therapy, preoperative tumor volume, length and maximum diameter (all P<0.05). Furthermore, multivariate Cox regression analysis showed that differentiation grade (HR=0.514, 95%CI: 0.366-0.723, P=0.019), TNM stage (HR=1.757, 95%CI: 1.267-2.612, P=0.015), adjuvant therapy (HR=0.669, 95%CI: 0.503-0.889, P=0.006), preoperative tumor volume (set <32 cm 3 as the dummy variable, 32-72 cm 3: HR=3.689, 95%CI: 2.415-5.637, P<0.001; >72 cm 3: HR=5.720, 95%CI: 3.606-9.075, P<0.001) were independent risk factors for OS. Finally, the C-index of OS by nomogram incorporated the statistically significant clinicopathological parameters was predicted to be 0.722 (95%CI: 0.687-0.757), which was significantly higher than the 7th AJCC TNM stage, the C-index 0.633 (95%CI: 0.595-0.671). In addition, the calibration curve of nomogram model was highly consistent with actual observation for the five-year OS rate, and the decision curve analysis also showed that nomogram model had higher clinical application potentials than TNM staging model in predicting survival prognosis of thoracic ESCC after surgery. Conclusion The nomogram incorporated preoperative tumor volume is of great value in predicting survival prognosis of patients with thoracic ESCC.

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    Expression and enrichment analysis of HOXC8 in esophageal cancer
    Zhang Mingxin, Lu Ning, Yao Li, Yuan Hui, Zhang Lingmin
    2020, 47 (5):  284-288.  doi: 10.3760/cma.j.cn371439-20190923-00019
    Abstract ( 583 )   HTML ( 25 )   PDF (3067KB) ( 235 )   Save

    Objective To investigate the expression of HOXC8 in esophageal cancer and its possible signaling pathway. Methods The RNA-Seq data of mRNA expression and clinical prognosis data of esophageal cancer dataset were downloaded and preprocessed from the TCGA (The Cancer Genome Atlas) database. The differentially expressed genes were analyzed, and the volcano map and heat map were drawn to visualize the screened differentially expressed genes. The patients with esophageal cancer were divided into high expression group and low expression group based on the median of HOXC8 expression, and survival analysis was performed using Kaplan-Meier method. GSEA 4.0.1 software was used for gene set enrichment analysis, and graphic analysis of multi-GSEA enrichment analysis was performed at the same time. Results After differential expression analysis of mRNA expression data of 161 esophageal cancer tissues and 11 paracancerous tissues, 3 454 differentially expressed genes were screened, including 2 317 up-regulated genes and 1 137 down-regulated genes. The results of cluster analysis showed that differential expression can effectively distinguish esophageal cancer from adjacent tissues, indicating that the above differential expression results had good accuracy. Difference analysis and paired difference analysis showed that HOXC8 was significantly overexpressed in esophageal cancer, and the differences with tissues adjacent to cancer were statistically significant (t=5.333, P<0.001; t=3.101, P=0.007). After removing samples with a survival time of less than 30 days, a total of 107 samples were used. The results showed that patients with high expression of HOXC8 (n=54) had a worse prognosis, with a median survival time of 553 days (95%CI: 396-710), and the median survival time of patients with low expression of HOXC8 (n=53) was 784 days (95%CI: 62-1 506), with a statistically significant difference (χ 2=4.153, P=0.042), suggesting that HOXC8 was an oncogene. The results of GSEA analysis showed that the samples with high expression of HOXC8 enriched the cell cycle, spliceosome and other related gene sets, while the samples with low expression of HOXC8 enriched the phosphatidylinositol signaling pathway and other related gene sets. Conclusion HOXC8 is significantly overexpressed in esophageal cancer, and patients with high expression of HOXC8 have a worse prognosis. It may regulate the occurrence and development of esophageal cancer through the involvement of cell cycle, spliceosome, phosphatidylinositol signaling pathway and other signaling pathways.

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    Reviews
    Research progress of Bub1 in cancers
    Wang Mengni, Lyu Weiguo
    2020, 47 (5):  289-292.  doi: 10.3760/cma.j.cn371439-20200113-00020
    Abstract ( 803 )   HTML ( 41 )   PDF (660KB) ( 453 )   Save

    Bub1 is essential for assembling of the functional spindle assembly checkpoint (SAC) to guarantee the correct separation of sister chromatids. Abnormal expression of Bub1 can elevate defects in SAC function, chromosome instability, and the incidence of aneuploidy. Several recent studies indicate that aberrantly expressed Bub1 may promote the tumorigenesis via regulating cell proliferation, invasion, migration and cancer stem cell activation. Further understanding of the mechanism of Bub1 in malignancy may provide new targeted therapy strategies.

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    Mining and prediction of glioma-related genes
    Zhao Congxuan, Yu Tao
    2020, 47 (5):  293-296.  doi: 10.3760/cma.j.cn371439-20200123-00021
    Abstract ( 683 )   HTML ( 15 )   PDF (659KB) ( 266 )   Save

    Glioma is the most common primary tumor of the central nervous system. Although clinicians select multimodal treatment, the overall prognosis of most patients is still very poor, especially glioblastoma (GB), which is related to the biological characteristics of glioma. Genomic alterations have been confirmed to be closely related to the tumorigenesis of GB. Different gene expression has a guiding therapeutic and prognostic predictive function for gliomas, so the deeper understanding of the relationship between genes and diseases behind gliomas will enable us to explore potential individualized targeted therapies for patients.

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    High mobility group protein B1 and breast cancer
    Zhang Yanqiang, Zhang Yang, Li Chunhua, Zhang Dianping, Liu Baoguo, Peng Xiangeng
    2020, 47 (5):  297-300.  doi: 10.3760/cma.j.cn371439-20200414-00022
    Abstract ( 508 )   HTML ( 29 )   PDF (658KB) ( 230 )   Save

    High mobility group protein B1 (HMGB1) is a kind of nuclear protein widely existing in cells, which is released or secreted from cells by stress in the body and plays a key role in the survival or death pathways of cells. HMGB1 has a huge biological function and is the main regulator of major diseases such as inflammatory diseases and tumors. HMGB1 is closely related to the proliferation, differentiation, migration, apoptosis and drug resistance of tumor cells. With the continuous deepening of research on HMGB1, it is found that HMGB1 plays an important role in the occurrence, development, metastasis and drug resistance of breast cancer. Combined with the research status of HMGB1, the expression of HMGB1 in breast cancer is discussed to provide a new therapeutic scheme for clinical treatment.

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    A new predictor of the efficacy of immunotherapy for non-small cell lung cancer—lung immune prognostic index (LIPI)
    Zhang Aixia, Sun Yahong
    2020, 47 (5):  301-303.  doi: 10.3760/cma.j.cn371439-20190926-00023
    Abstract ( 1200 )   HTML ( 44 )   PDF (653KB) ( 505 )   Save

    Immune checkpoint inhibitors have demonstrated the ability to significantly improve survival across a range of cancers. So predictive markers are required to guide treatment decisions. The lung immune prognostic index (LIPI) is recently developed to predict immune checkpoint inhibitors (ICIs) treatment outcomes for non-small cell lung cancer. The pre-treatment LIPI is a convenient prognostic marker able to idenjpgy ICIs-treated patient groups with significantly different survival and response outcomes.

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    Mechanism of long non-coding RNA GHET1 in tumors of the digestive system
    Ma Yingji, Sun Libin, Qiu Wensheng
    2020, 47 (5):  304-307.  doi: 10.3760/cma.j.cn371439-20200209-00024
    Abstract ( 467 )   HTML ( 21 )   PDF (659KB) ( 238 )   Save

    Gastric cancer highly expressed transcript 1 (GHET1) is first found in gastric cancer and is a long non-coding RNA (lncRNA). GHET1 is located on chromosome 7q36.1, and is highly expressed in many tumors. High expression of GHET1 is closely related to poor prognosis. Studies have found that GHET1 is involved in regulating many physiological and pathological processes of the body through interaction with microRNAs (miRNAs) or proteins, especially in digestive system tumors, and is expected to become a valuable tumor marker and therapeutic target in the future.

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    Molecular mechanisms of cetuximab resistance in metastatic colorectal cancer
    Su Hao, Liu Wenjie, Bao Mandula, Luo Shou, Wang Xuewei, Zhao Chuanduo, Liu Qian, Wang Xishan, Zhou Zhixiang, Zhou Haitao
    2020, 47 (5):  308-311.  doi: 10.3760/cma.j.cn371439-20200208-00025
    Abstract ( 1102 )   HTML ( 40 )   PDF (659KB) ( 531 )   Save

    Cetuximab has become an important molecular targeted drug for the treatment of metastatic colorectal cancer (mCRC), which increases the curative effect of chemotherapy and prolongs the survival time. However, some patients develop insensitiveness or resistance to cetuximab, while the complicated molecular mechanisms are not quite clear. With the deep research in epidermal growth factor receptor (EGFR) signaling pathway, the genetic alteration of KRAS, BRAF, PTEN and PIK3CA and polymorphism of microRNA (miRNA) have been proved to associated with cetuximab resistance. Wnt signaling pathway with its negative regulator RNF43 is also considered to be related with cetuximab resistance in recent studies. The review of the progress on molecular mechanisms of cetuximab resistance in mCRC can establish theoretical basis for finding out reasonable drugs to overcome the resistance.

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    Research progress of PD-1/PD-L1 inhibitors in ovarian cancer
    Xue Chen, Zhao Yue, Shi Guang, Tang Yan
    2020, 47 (5):  312-315.  doi: 10.3760/cma.j.cn371439-20200321-00026
    Abstract ( 917 )   HTML ( 32 )   PDF (662KB) ( 547 )   Save

    Ovarian cancer has the characteristics of difficulty in early diagnosis, easy recurrence and resistance, so its mortality rate ranks first in gynecological tumors. Existing treatment methods such as surgery, radiotherapy and chemotherapy are not very effective, thus immunotherapy has emerged. Immune checkpoint programmed death-1 (PD-1) and its ligand PD-L1 can inhibit T cell proliferation and mediate tumor immune escape. At present, PD-1/PD-L1 inhibitors have made breakthrough progress in a variety of solid tumors, and they have achieved initial efficacy in clinical treatment of ovarian cancer.

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