血清細(xì)胞因子和microRNA檢測在宮頸疾病中的臨床應(yīng)用價(jià)值
發(fā)布時(shí)間：2018-08-16 07:36
【摘要】：研究背景：宮頸癌是全球女性第三大惡性腫瘤，早期診斷宮頸癌及癌前病變是降低宮頸癌發(fā)生率和死亡率的重要方法。循環(huán)炎癥標(biāo)志物和microRNA有可能成為宮頸癌相關(guān)的生物標(biāo)志物用于腫瘤的早期檢測或預(yù)后。研究目的：研究宮頸癌、宮頸疾病和健康對照的血清細(xì)胞因子和miRNA譜的差異，篩選新的標(biāo)記物，并建立具有高靈敏度和特異度的宮頸癌輔助診斷模型。方法：從2010年1月至2010年12月收集原發(fā)性宮頸癌95例、宮頸病變80例及健康對照組85例血清標(biāo)本，用Luminex200檢測10項(xiàng)血清細(xì)胞因子水平，生物信息學(xué)分析三組間腫瘤標(biāo)志物和細(xì)胞因子的表達(dá)差異，建立輔助診斷模型，并用驗(yàn)證集進(jìn)行驗(yàn)證；對其中60名宮頸癌患者進(jìn)行為期2年的隨訪，統(tǒng)計(jì)分析她們的2年總生存率及影響預(yù)后的因素，建立預(yù)后模型；2012年3月至2013年5月收集133例宮頸癌患者、123例宮頸病變患者和106例健康對照的血清標(biāo)本，每組各由21例血清標(biāo)本組成3個(gè)血清池，共組成9個(gè)血清池進(jìn)行Solexa測序，篩選并預(yù)測新的miRNA，對預(yù)測的新miRNA用實(shí)時(shí)定量反轉(zhuǎn)錄聚合酶鏈反應(yīng)（Quantitative real-time polymerase chain reaction）進(jìn)行驗(yàn)證，并用Receiveroperating characteristic（ROC）曲線分析新miRNA對宮頸疾病的輔助診斷能力。 結(jié)果：（1）在用單一炎性細(xì)胞因子和腫瘤標(biāo)志物區(qū)分宮頸疾病組和健康對照組時(shí)，interleukin(IL)-8的Area under curve (AUC)最大為0.921(95%：0.885-0.958)，在24.04pg/ml的臨界值時(shí)，其敏感度為84.8％，特異度為88.1％，高于SCC；用二元logistic回歸建立的多參數(shù)診斷模型模(包括Squamous cell carcinoma associatedantigen （SCC）、IL-8和Monocyte chemoattractant protein1（MCP-1)）的AUC為0.939。鑒別診斷宮頸癌和宮頸病變時(shí)，單個(gè)腫瘤標(biāo)志物和細(xì)胞因子的價(jià)值均較低，運(yùn)用分類樹多參數(shù)分析建立的模型（包括SCC和IL-10）的ROC曲線下為0.729，較臨床上常用的SCC有了一定提高。（2）對60例I-II期宮頸癌患者的2年隨訪表明，術(shù)前血清Carbohydrate antigen153（CA153）、SCC、IL-10和TNF-α（tumornecrosis factor-α）水平與宮頸癌患者的兩年總生存率相關(guān)。COX多參數(shù)分析顯示CA153≥17.60U/ml、SCC≥1.60ng/ml和TNF-α≥10.60pg/ml可能為宮頸癌I-II期患者的獨(dú)立預(yù)后危險(xiǎn)因素。依據(jù)血清CA153水平和TNF α水平可以對宮頸癌患者進(jìn)行危險(xiǎn)分層，分為三層：①血清CA153≥17.60μg/L的水平；②血清CA15317.60μg/L和TNFα≥10.60pg/ml；③血清CA15317.60μg/L和TNF α10.60pg/ml。這三組間的兩年總生存率分別為33.3％、60.0％和93.9％。（3）通過對宮頸癌患者血清池進(jìn)行Solexa測序，，我們預(yù)測、篩選和驗(yàn)證了5個(gè)新的miRNA。在區(qū)分宮頸疾病和健康對照時(shí)，PmiR-3的AUC為0.924(95％CI:0.888-0.959)，當(dāng)約登指數(shù)最大時(shí)，其敏感度為92.7％，特異性為78.3％；鑒別診斷宮頸癌與宮頸病變患者時(shí)，PmiR-1的AUC達(dá)到了0.893(95％CI：0.842-0.943)，約登指數(shù)最大時(shí)的敏感度為85.9％，特異性為83.9％。文章已經(jīng)提交，尚未接收，所以以上新miRNA為暫命名，待文章接收后，miRbase會對新miRNA進(jìn)行命名。 結(jié)論：運(yùn)用多元統(tǒng)計(jì)分析方法，將腫瘤標(biāo)志物和炎性細(xì)胞因子聯(lián)合運(yùn)用可以提高其輔助診斷宮頸疾病的能力，IL-8、IL-10和MCP-1可以為宮頸癌的早期預(yù)警提供重要參考價(jià)值。炎性細(xì)胞因子TNF α在宮頸癌的預(yù)后中也發(fā)揮重要作用。循環(huán)血清miRNAs有望成為宮頸癌相關(guān)的生物標(biāo)志物的用于疾病的早期診斷，并為宮頸癌的機(jī)理研究提供新的方向。
[Abstract]:Background: Cervical cancer is the third largest malignant tumor in the world. Early diagnosis of cervical cancer and precancerous lesions is an important method to reduce the incidence and mortality of cervical cancer. Methods: From January 2010 to December 2010, 95 cases of primary cervical cancer, 80 cases of cervical lesions and 85 cases of healthy control group were collected and tested by Luminex 200. The levels of 10 serum cytokines were measured, and the expression of tumor markers and cytokines were analyzed by bioinformatics. The adjuvant diagnosis model was established and validated by validation set. Sixty patients with cervical cancer were followed up for 2 years. Their 2-year overall survival rate and prognostic factors were analyzed and prognostic models were established. From March 2012 to May 2013, serum samples from 133 patients with cervical cancer, 123 patients with cervical lesions and 106 healthy controls were collected. Each group consisted of 21 serum samples and consisted of 3 serum pools. Solexa sequencing was performed in 9 serum pools to screen and predict new microRNAs. The predicted new microRNAs were identified by real-time quantitative reverse transcription polymerase chain reaction (Qu-PCR). Antitative real-time polymerase chain reaction was used to validate the method and the Receiver operating characteristic (ROC) curve was used to analyze the auxiliary diagnostic ability of new microRNAs for cervical diseases.
Results: (1) The area under curve (AUC) of interleukin (IL) - 8 was 0.921 (95%:0.885-0.958) when using a single inflammatory cytokine and tumor marker to distinguish cervical disease group from healthy control group, and the sensitivity and specificity were 84.8% and 88.1% at the critical value of 24.04 pg/ml, which were higher than those of SCC. The AUC of the parametric diagnostic model (including Squamous cell carcinoma associatedantigen (SCC), IL-8 and Monocyte chemoattractant protein 1 (MCP-1)) was 0.939. In the differential diagnosis of cervical cancer and cervical lesions, the value of single tumor marker and cytokines was low. The model established by using classification tree multiparametric analysis (including SCC and IL-10) The two-year follow-up of 60 patients with stage I-II cervical cancer showed that preoperative serum levels of Carbohydrate antigen 153 (CA153), SCC, IL-10 and TNF-alpha were correlated with the two-year overall survival rate of patients with cervical cancer. SCC (> 1.60ng/ml) and TNF-a (> 10.60pg/ml) may be independent prognostic factors for patients with stage I-II cervical cancer. According to serum CA153 level and TNF-a level, risk stratification of patients with cervical cancer can be divided into three layers: serum CA153 (> 17.60ug/L); serum CA15317.60ug/L and TNF-a (> 10.60pg/ml); serum CA15317.60 pg/ml; and TNF-a (> 10.60pg/ml The two-year overall survival rates of the three groups were 33.3%, 60.0% and 93.9%. (3) By sequencing the serum pools of cervical cancer patients with Solexa, we predicted that five new microRNAs were screened and validated. The sensitivity and specificity were 92.7% and 78.3% respectively, and the AUC of PmiR-1 was 0.893 (95% CI: 0.842-0.943) and 85.9% and 83.9% respectively in the differential diagnosis of cervical cancer and cervical lesions. E will name the new miRNA.
Conclusion: Combined use of tumor markers and inflammatory cytokines can improve the ability of assistant diagnosis of cervical diseases. IL-8, IL-10 and MCP-1 can provide important reference value for early warning of cervical cancer. Inflammatory cytokine TNFalpha also plays an important role in the prognosis of cervical cancer. IRNAs are expected to become biomarkers related to cervical cancer for the early diagnosis of diseases, and provide a new direction for the study of the mechanism of cervical cancer.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R737.33
[Abstract]:Background: Cervical cancer is the third largest malignant tumor in the world. Early diagnosis of cervical cancer and precancerous lesions is an important method to reduce the incidence and mortality of cervical cancer. Methods: From January 2010 to December 2010, 95 cases of primary cervical cancer, 80 cases of cervical lesions and 85 cases of healthy control group were collected and tested by Luminex 200. The levels of 10 serum cytokines were measured, and the expression of tumor markers and cytokines were analyzed by bioinformatics. The adjuvant diagnosis model was established and validated by validation set. Sixty patients with cervical cancer were followed up for 2 years. Their 2-year overall survival rate and prognostic factors were analyzed and prognostic models were established. From March 2012 to May 2013, serum samples from 133 patients with cervical cancer, 123 patients with cervical lesions and 106 healthy controls were collected. Each group consisted of 21 serum samples and consisted of 3 serum pools. Solexa sequencing was performed in 9 serum pools to screen and predict new microRNAs. The predicted new microRNAs were identified by real-time quantitative reverse transcription polymerase chain reaction (Qu-PCR). Antitative real-time polymerase chain reaction was used to validate the method and the Receiver operating characteristic (ROC) curve was used to analyze the auxiliary diagnostic ability of new microRNAs for cervical diseases.
Results: (1) The area under curve (AUC) of interleukin (IL) - 8 was 0.921 (95%:0.885-0.958) when using a single inflammatory cytokine and tumor marker to distinguish cervical disease group from healthy control group, and the sensitivity and specificity were 84.8% and 88.1% at the critical value of 24.04 pg/ml, which were higher than those of SCC. The AUC of the parametric diagnostic model (including Squamous cell carcinoma associatedantigen (SCC), IL-8 and Monocyte chemoattractant protein 1 (MCP-1)) was 0.939. In the differential diagnosis of cervical cancer and cervical lesions, the value of single tumor marker and cytokines was low. The model established by using classification tree multiparametric analysis (including SCC and IL-10) The two-year follow-up of 60 patients with stage I-II cervical cancer showed that preoperative serum levels of Carbohydrate antigen 153 (CA153), SCC, IL-10 and TNF-alpha were correlated with the two-year overall survival rate of patients with cervical cancer. SCC (> 1.60ng/ml) and TNF-a (> 10.60pg/ml) may be independent prognostic factors for patients with stage I-II cervical cancer. According to serum CA153 level and TNF-a level, risk stratification of patients with cervical cancer can be divided into three layers: serum CA153 (> 17.60ug/L); serum CA15317.60ug/L and TNF-a (> 10.60pg/ml); serum CA15317.60 pg/ml; and TNF-a (> 10.60pg/ml The two-year overall survival rates of the three groups were 33.3%, 60.0% and 93.9%. (3) By sequencing the serum pools of cervical cancer patients with Solexa, we predicted that five new microRNAs were screened and validated. The sensitivity and specificity were 92.7% and 78.3% respectively, and the AUC of PmiR-1 was 0.893 (95% CI: 0.842-0.943) and 85.9% and 83.9% respectively in the differential diagnosis of cervical cancer and cervical lesions. E will name the new miRNA.
Conclusion: Combined use of tumor markers and inflammatory cytokines can improve the ability of assistant diagnosis of cervical diseases. IL-8, IL-10 and MCP-1 can provide important reference value for early warning of cervical cancer. Inflammatory cytokine TNFalpha also plays an important role in the prognosis of cervical cancer. IRNAs are expected to become biomarkers related to cervical cancer for the early diagnosis of diseases, and provide a new direction for the study of the mechanism of cervical cancer.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R737.33
【共引文獻(xiàn)】
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