CAJ | 학술논문

目的：探讨抗菌肽hCAP18／LL?37在卵巢癌微环境中的作用及表达调控机制。方法采用侵袭实验检测巨噬细胞对卵巢癌细胞SKOV3侵袭能力的影响。采用实时荧光定量PCR（ qRT?PCR）和Western blot检测hCAP18／LL?37和versican V1蛋白的表达。采用干扰质粒抑制 SKOV3细胞中versican V1的表达，分析巨噬细胞hCAP18／LL?37的表达及SKOV3细胞的侵袭能力。结果共培养组（SKOV3细胞与巨噬细胞共培养）的侵袭穿膜数为（112．8±17．1）个，高于SKOV3培养组[SKOV3细胞单独培养，（8．2±1．9）个]，差异有统计学意义（P＜0．05）。 hCAP18／LL?37中和抗体共培养组（SKOV3细胞、巨噬细胞和hCAP18／LL?37中和抗体共培养）的侵袭穿膜细胞数为（22．2±5．6）个，少于对照IgG共培养组[SKOV3细胞、巨噬细胞和对照IgG共培养，（100．6±25．2）个]，差异有统计学意义（P＜0．05）。与SKOV3细胞共培养后，巨噬细胞中 hCAP18／LL?37蛋白和 mRNA水平均升高，而在SKOV3细胞中无变化。与巨噬细胞共培养后，卵巢癌细胞中versican V1蛋白的表达和分泌均升高。干扰卵巢癌SKOV3细胞中versican V1的表达（ SKOV3ver－／－细胞）后，巨噬细胞中hCAP18／LL?37蛋白和mRNA水平均低于对照细胞株（ SKOV3ver＋／＋细胞）；与巨噬细胞共培养后，SKOV3ver－／－细胞的穿膜细胞数[（24．8±4．6）个]低于SKOV3ver＋／＋细胞[（104．6±16．0）个]，差异有统计学意义（P＜0．05）。结论卵巢癌微环境中，巨噬细胞表达分泌的抗菌肽hCAP18／LL?37促进卵巢癌细胞的侵袭，其表达受到肿瘤细胞分泌的versicanV1蛋白调控。
목적：탐토항균태hCAP18／LL?37재란소암미배경중적작용급표체조공궤제。방법채용침습실험검측거서세포대란소암세포SKOV3침습능력적영향。채용실시형광정량PCR（ qRT?PCR）화Western blot검측hCAP18／LL?37화versican V1단백적표체。채용간우질립억제 SKOV3세포중versican V1적표체，분석거서세포hCAP18／LL?37적표체급SKOV3세포적침습능력。결과공배양조（SKOV3세포여거서세포공배양）적침습천막수위（112．8±17．1）개，고우SKOV3배양조[SKOV3세포단독배양，（8．2±1．9）개]，차이유통계학의의（P＜0．05）。 hCAP18／LL?37중화항체공배양조（SKOV3세포、거서세포화hCAP18／LL?37중화항체공배양）적침습천막세포수위（22．2±5．6）개，소우대조IgG공배양조[SKOV3세포、거서세포화대조IgG공배양，（100．6±25．2）개]，차이유통계학의의（P＜0．05）。여SKOV3세포공배양후，거서세포중 hCAP18／LL?37단백화 mRNA수평균승고，이재SKOV3세포중무변화。여거서세포공배양후，란소암세포중versican V1단백적표체화분비균승고。간우란소암SKOV3세포중versican V1적표체（ SKOV3ver－／－세포）후，거서세포중hCAP18／LL?37단백화mRNA수평균저우대조세포주（ SKOV3ver＋／＋세포）；여거서세포공배양후，SKOV3ver－／－세포적천막세포수[（24．8±4．6）개]저우SKOV3ver＋／＋세포[（104．6±16．0）개]，차이유통계학의의（P＜0．05）。결론란소암미배경중，거서세포표체분비적항균태hCAP18／LL?37촉진란소암세포적침습，기표체수도종류세포분비적versicanV1단백조공。
Objective To investigate the effect of antibacterial peptide hCAP18/LL?37 on ovarian cancer microenvironment and the regulatory mechanism of its expression. Methods We assessed the effect of macrophage?promoted ovarian cancer cells invasion using BioCoat Matrigel invasion chamber. The expressions of hCAP18/LL?37 and versican V1 were determined by real?time PCR and Western blot analysis. SKOV3 cells were transfected with shRNA plasmid to abrogate the expression of versican V1, and then the expression of hCAP18/LL?37 in macrophages and the invasiveness of SKOV3 cells were assayed. Results The Matrigel invasion assay showed that after co?culture with macrophages for 4 days,the number of penetrated SKOV3 cells was 112. 8 ± 17. 1/per high power field, significantly higher than that in the SKOV3 cells cultured alone (8.2±1.9/per high power field) (P<0.05). Addition of hCAP/LL?37 neutralizing antibody into the co?cultured macrophage?SKOV3 cells markedly inhibited the macrophage?promoted SKOV3 cells invasion. The penetrated SKOV3 cells was 22.2±5.6/per high power field, significantly lower than the 100.6± 25.2/per high power field in the control macrophage?SKOV3 co?cultured cells (P<0.05). The expressions of hCAP18/LL?37 mRNA and protein in macrophages were remarkably enhanced upon co?culture with SKOV3 cells, but not changed in SKOV3 cells cultured alone. The expression and secretion of versican V1 in the ovarian cancer cells were also significantly increased after co?cultured with macrophages. Knockdown of versican V1 in SKOV3 cells by small interfering RNA significantly reduced the expression of hCAP18/LL?37 <br> mRNA and protein in the macrophages, as well as decreased the invasiveness of SKOV3 cells (P<0.05). Conclusions In the cancer microenvironment, the macrophage?secreted hCAP18/LL?37 promote the invasiveness of ovarian cancer cells, and the hCAP18/LL?37 expression is regulated by versican V1 protein released by ovarian cancer cells.