Zhao Yi Ming, Yang Gang, You Lei, Hu Ya, Zhao Yu Pei
Department of General Surgery,PUMC Hospital,CAMS and PUMC,Beijing 100730,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2020 Feb 28;42(1):7-15. doi: 10.3881/j.issn.1000-503X.11192.
To unravel the role of hematopoietic pre-B-cell leukemia transcription factor interacting protein(HPIP)in the proliferation,cell cycle,and apoptosis of pancreatic ductal adenocarcinoma(PDAC)cells. The HPIP expression in PDAC tissue was determined by immunohistochemical staining.Knockdown of HPIP was accomplished in MIA PaCa-2 and BxPC-3 cell lines by transient transfection of HPIP siRNA and validated by Western blotting.Cell proliferation was assessed using the cell counting kit-8 assay and colony formation assay.Cell cycle and apoptosis were detected by flow cytometry.Western blotting was performed to detect the expression levels of cyclin D1,caspase 7,and cleaved caspase 7. HPIP was overexpressed in PDAC tissue compared with matched adjacent pancreatic tissue(=-2.060,=0.039).Knockdown of HPIP inhibited the proliferation of MIA PaCa-2 and BxPC-3 cells(all <0.05).Knockdown of HPIP significantly reduced the positive colonies formed by MIA PaCa-2 and BxPC-3 cells(=4.706,=0.009;=9.514,=0.000).Knockdown of HPIP decreased the proportion of S phase cells(=7.642,=0.001;=2.714,=0.051)and increased the proportion of G/G phase cells(=3.244,=0.031;=6.095,=0.003)in MIA PaCa-2 and BxPC-3 cells.Meanwhile,knockdown of HPIP increased the proportions of late-phase MIA PaCa-2 and BxPC-3 cells(=24.58,=0.000;=36.45,=0.000)and the overall apoptosis rate(=29.43,=0.000;=43.52,=0.000).In MIA PaCa-2 and BxPC-3 cells,knockdown of HPIP decreased the expression level of cyclin D1(=6.705,=0.002;=6.238,=0.003)and increased the expression level of cleaved caspase 7(=3.991,=0.016;=6.536,=0.002). HPIP is overexpressed in PDAC tissue.Knockdown of HPIP inhibits the proliferation and G/G to S transition of PDAC cells.Meanwhile,knockdown of HPIP promotes the apoptosis of PDAC cells.Thus,HPIP may act as an oncogene in PDAC.
为了阐明造血前B细胞白血病转录因子相互作用蛋白（HPIP）在胰腺导管腺癌（PDAC）细胞增殖、细胞周期及凋亡中的作用。通过免疫组织化学染色检测PDAC组织中HPIP的表达。在MIA PaCa-2和BxPC-3细胞系中通过瞬时转染HPIP siRNA实现HPIP基因敲低，并通过蛋白质免疫印迹法进行验证。使用细胞计数试剂盒-8法和集落形成试验评估细胞增殖。通过流式细胞术检测细胞周期和凋亡情况。采用蛋白质免疫印迹法检测细胞周期蛋白D1、半胱天冬酶7及裂解的半胱天冬酶7的表达水平。与配对的相邻胰腺组织相比，HPIP在PDAC组织中高表达（=-2.060，=0.039）。敲低HPIP可抑制MIA PaCa-2和BxPC-3细胞的增殖（均<0.05）。敲低HPIP显著减少了MIA PaCa-2和BxPC-3细胞形成的阳性集落（=4.706，=0.009；=9.514，=0.000）。敲低HPIP降低了MIA PaCa-2和BxPC-3细胞中S期细胞的比例（=7.642，=0.001；=2.714，=0.051），并增加了G/G期细胞的比例（=3.244，=0.031；=6.095，=0.003）。同时，敲低HPIP增加了MIA PaCa-2和BxPC-3细胞晚期细胞的比例（=24.58，=0.000；=36.45，=0.000）及总体凋亡率（=29.43，=0.000；=43.52，=0.000）。在MIA PaCa-2和BxPC-3细胞中，敲低HPIP降低了细胞周期蛋白D1的表达水平（=6.705，=0.002；=6.238，=0.003），并增加了裂解的半胱天冬酶7的表达水平（=3.991，=0.016；=6.536，=0.002）。HPIP在PDAC组织中高表达。敲低HPIP可抑制PDAC细胞的增殖及G/G期向S期的转变。同时，敲低HPIP可促进PDAC细胞的凋亡。因此，HPIP可能在PDAC中作为癌基因发挥作用。
