氢气对原代人大肠腺癌细胞生长的抑制作用及其影响因素

Inhibitory effect of hydrogen on growth of primary human colorectal adenocarcinoma cells and its influencing factors

目的探讨氢气对原代人大肠腺癌细胞生长的抑制作用及其影响因素。方法取30例大肠腺癌患者经手术切除的肿瘤组织标本,经胰蛋白酶消化后进行原代细胞培养。将肿瘤细胞分为对照组、氢气组,对照组给予含10%血清的DMEM/F12培养液培养,氢气组给予含10%血清饱和氢气(≥0.6 mmol/L)的DMEM/F12培养液培养。采用MTT法检测两组细胞活性(OD值),克隆形成实验检测细胞增殖情况,流式细胞仪检测细胞凋亡率。收集患者的临床病理资料,分析可能影响氢气抗肿瘤作用的因素。结果氢气组15例患者的原代结肠癌细胞OD值、形成的克隆数量均低于对照组(P均<0.05),细胞凋亡率高于对照组(P<0.05),提示氢气干预有效。对15例氢气干预有效的大肠腺癌原代细胞来源患者的临床病理参数进行单因素分析结果显示,血浆丙二醛(MDA)>3μmol/L和癌细胞Ki67表达阳性率>70%患者的原代癌细胞被氢气抑制的概率高于血浆MDA≤3μmol/L和癌细胞Ki67表达阳性率≤70%患者(P均<0.05);患者性别、年龄、肿瘤位置及Dukes分期对原代癌细胞是否被氢气抑制无显著影响(P均>0.05)。结论氢气对原代人大肠腺癌细胞生长具有一定的抑制作用;血浆MDA含量和肿瘤组织Ki67表达可能影响氢气的临床抗肿瘤效果。

Objective To investigate the inhibitory effect of hydrogen on the growth of primary human colorectal adenocarcinoma cells and its influencing factors.Methods Resected tumor fragments from 30 cases of patients with colorectal adenocarcinoma were obtained and primarily cultured after trypsin digestion.Tumor cells were divided into the control group and hydrogen group.The cells in the control group were cultured with DMEM/F12 medium containing 10%serum,and the cells in the hydrogen group were cultured with DMEM/F12 medium containing 10%serum and saturated hydrogen(≥0.6 mmol/L).The cell viability(OD value)of the two groups was detected by MTT,the cell proliferation was assessed by cloning formation assay,and the apoptotic rate was measured by flow cytometry.The clinicopathological data of patients were collected to analyze the factors that might affect the anti-tumor effect of hydrogen.Results The OD values and formed clone number of primary colorectal cancer cells from 15 patients in the hydrogen group were lower(both P<0.05)and the apoptosis rate was higher than that of the control group(P<0.05),suggesting that hydrogen intervention was effective.Univariate analysis was carried out on the clinicopathological parameters of primary colorectal adenocarcinoma primary cells derived from 15 patients with effective hydrogen intervention.The results showed that the primary colorectal adenocarcinoma cells were more likely to be inhibited by hydrogen in patients with plasma malondialdehyde(MDA)>3μmol/L and positive rate of Ki67 expression of cancer cells>70%than in patients with plasma malondialdehyde(MDA)>3μmol/L and the positive rate of Ki67 expression of cancer cells>70%(P<0.05).The gender,age of patients,location and Dukes stage of tumors had no significant effect on whether primary cancer cells were inhibited by hydrogen(all P<0.05).Conclusion Hydrogen has a certain inhibitory effect on the growth of human colorectal adenocarcinoma cells,the plasma MDA content and the expression of Ki67 in the tumor tissues may affect the clinical antitumor effect of hydrogen.