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SPARCL1表达下调对胆囊癌吉西他滨耐药的作用机制及临床意义

和华 刘少朋 刘海潮 白明辉

和华, 刘少朋, 刘海潮, 白明辉. SPARCL1表达下调对胆囊癌吉西他滨耐药的作用机制及临床意义[J]. 中华全科医学, 2022, 20(10): 1666-1671. doi: 10.16766/j.cnki.issn.1674-4152.002674
引用本文: 和华, 刘少朋, 刘海潮, 白明辉. SPARCL1表达下调对胆囊癌吉西他滨耐药的作用机制及临床意义[J]. 中华全科医学, 2022, 20(10): 1666-1671. doi: 10.16766/j.cnki.issn.1674-4152.002674
HE Hua, LIU Shao-peng, LIU Hai-chao, BAI Ming-hui. The mechanism and clinical significance of SPARCL1 down-regulation expression on gemcitabine resistance in gallbladder cancer[J]. Chinese Journal of General Practice, 2022, 20(10): 1666-1671. doi: 10.16766/j.cnki.issn.1674-4152.002674
Citation: HE Hua, LIU Shao-peng, LIU Hai-chao, BAI Ming-hui. The mechanism and clinical significance of SPARCL1 down-regulation expression on gemcitabine resistance in gallbladder cancer[J]. Chinese Journal of General Practice, 2022, 20(10): 1666-1671. doi: 10.16766/j.cnki.issn.1674-4152.002674

SPARCL1表达下调对胆囊癌吉西他滨耐药的作用机制及临床意义

doi: 10.16766/j.cnki.issn.1674-4152.002674
基金项目: 

河南省卫生健康委员会联合共建项目 LHGJ20191194

洛阳市科技医疗卫生项目 1820003A

详细信息
    通讯作者:

    刘少朋, E-mail: 1281733563@qq.com

  • 中图分类号: R735.8  R730.43

The mechanism and clinical significance of SPARCL1 down-regulation expression on gemcitabine resistance in gallbladder cancer

  • 摘要:   目的   研究富含半胱氨酸酸性分泌型糖蛋白类似物1(SPARCL1)在胆囊癌中的作用。   方法   将人胆囊癌吉西他滨耐药细胞株GBC-SD/GEM分为空白对照组和SPARCL1过表达转染组。检测转染后SPARCL1表达、各细胞增殖能力及基质金属蛋白酶-9(MMP-9)、波形蛋白(VIM)、纤维连接蛋白1(FN1)表达;检测2014年1月—2017年1月在洛阳中心医院行手术治疗的50例胆囊癌(吉西他滨耐药组、敏感组)及癌旁组织中SPARCL1表达,分析其与患者临床病理特征及预后间的关系。   结果   转染组SPARCL1基因相对表达量(29.12±1.10)及蛋白相对表达量(23.08±2.15)均显著高于空白对照组(3.34±0.98、2.58±0.71,均P<0.05)。转染组细胞增殖数及MMP-9、VIM、FN1相对表达量分别为33.09±12.11、1.69±0.75、1.78±0.43、1.62±0.31,均低于对照组(327.15±9.28、3.81±0.78、4.12±0.24、4.69±0.63,均P<0.05)。耐药组、敏感组及癌旁组织中SPARCL1表达量分别为5.65±2.01、15.02±1.17、28.46±2.53,组间差异有统计学意义(P<0.05)。SPARCL1表达与性别、年龄、病理类型、肿瘤大小、CA19-9、CEA和肿瘤分期无相关性(均P>0.05),而与淋巴结转移和肿瘤分化程度有相关性(均P<0.05)。耐药组患者中位生存期明显低于敏感组(14.77个月vs. 27.28个月)。   结论   SPARCL1在胆囊癌中低表达,且与吉西他滨化疗耐药及预后相关。

     

  • 图  1  SPARCL1对GBC-SD/GEM细胞克隆的影响(n=10)

    注:A为空白对照组; B为SPARCL1转染组; C为2组细胞克隆形成数比较, aP<0.01。

    Figure  1.  Effect of SPARCL1 on GBC-SD/GEM cell clones(n=10)

    图  2  转染后细胞内MMP-9、VIM、FN1表达

    注:A为蛋白印迹实验;B为2组蛋白相对表达量比较,aP<0.01。

    Figure  2.  Expression of MMP-9, VIM and FN1 in cells after transfection

    图  3  SPARCL1在胆囊癌组织及癌旁组织中的表达(HE染色,×200)

    注:A为敏感组,B为耐药组,C为癌旁正常组织。

    Figure  3.  Expression of SPARCL1 in gallbladder carcinoma tissues and adjacent tissues(HE dyed, ×200)

    图  4  吉西他滨耐药组患者与敏感组患者的生存期比较(n=50)

    Figure  4.  Comparison of survival time of patients in gemcitabine resistant group and sensitive group(n=50)

    表  1  实时免疫荧光引物序列

    Table  1.   Real-time immunofluorescence primer sequence

    基因 引物序列
    SPARCL1 上游:TTGGCTCCTGGTGTTAGTTC
    下游:ATCCTGCTCTTGGTTTCCTT
    内参GADPH 上游:TGACTTCAACAGCGACACCCA
    下游:CACCCTGTTGCTGTAGCCAAA
    下载: 导出CSV

    表  2  不同组别中SPARCL1的表达(例)

    Table  2.   Expression of SPARCL1 in different groups(cases)

    组别 例数 SPARCL1
    阳性 阴性
    耐药组 29 3 26
    敏感组 21 10 11
    癌旁正常组织 50 43 7
    注:3组比较,χ2=43.394,P<0.001。
    下载: 导出CSV

    表  3  SPARCL1表达与胆囊癌临床病理因素的相关性(例)

    Table  3.   Correlation between SPARCL1 expression and clinicopathological factors in gallbladder carcinoma(cases)

    临床病理因素 类别 例数 SPARCL1 χ2 P
    阳性 阴性
    性别 男性 27 7 20 <0.001 0.990
    女性 23 6 17
    年龄(岁)[11] ≥55 17 3 14 0.934 0.334
    <55 33 10 23
    肿瘤直径(cm)[12] ≥2 9 2 7 0.081 0.775
    <2 41 11 30
    Nevin分期(Ⅱ~Ⅳ)[13] Ⅱ、Ⅲ 43 12 31 0.581 0.446
    7 1 6
    病理类型 腺癌 41 12 29 1.265 0.261
    腺鳞癌 9 1 8
    CA19-9(kU/L) ≥37 15 4 11 0.005 0.944
    <37 35 9 26
    CEA(μg/L) ≥5 28 8 20 0.219 0.640
    <5 22 5 17
    分化程度 高分化 22 12 10 16.638 <0.001
    中、低分化 28 1 27
    淋巴结转移 15 1 14 4.163 0.041
    35 12 23
    下载: 导出CSV

    表  4  胆囊癌预后的Cox多因素分析结果

    Table  4.   Cox multivariate analysis of prognosis of gallbladder carcinoma

    变量 例数 OS(月) B SE Wald χ2 P HR 95% CI
    有淋巴结转移 15 13 0.524 1.833 6.661 0.005 0.493 0.231~0.882
    高分化 22 23 0.924 0.085 0.285 0.697 1.272 0.688~3.101
    SPARCL1阳性表达 23 34 0.515 0.659 7.522 0.001 0.619 0.313~0.899
    注:变量赋值方法如下,淋巴结转移,1=有转移,2=无转移;分化程度,1=高分化,2=中低分化;SPARCL1,1=阳性,2=阴性。OS为总生存期(overall survival)。
    下载: 导出CSV
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  • 收稿日期:  2021-05-15
  • 网络出版日期:  2022-11-30

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