益气除痰方调控内质网应激反应抑制H1650肺癌生长的研究

Study of Yiqi Chutan Recipe in Suppressing H1650 Lung Cancer by Regulating Endoplasmic Reticulum Stress Response

【目的】探讨益气除痰方对肺腺癌H1650细胞的抑瘤作用及机制。【方法】细胞实验采用流式细胞术测定益气除痰方含药血清对H1650细胞凋亡的影响,Western blot法检测益气除痰方含药血清对H1650细胞生长阻滞和DNA损害诱导基因153(CHOP/GADD153)、生长阻滞和DNA损害诱导基因34(GADD34)表达的影响。动物实验通过复制BALB/C裸鼠H1650肺癌种植瘤模型,随机分为模型组、中药低剂量组(剂量为9.1 g·kg-1·d-1)、中药高剂量组(剂量为27.3 g·kg-1·d-1),测定各组瘤体体积与质量,Western blot法检测各组肺癌组织CHOP/GADD153、GADD34蛋白的表达。【结果】体积分数15%、30%含药血清组H1650细胞的凋亡率分别为(8.14±1.10)%、(18.21±3.07)%(P<0.05),30%含药血清组CHOP/GADD153、GADD34蛋白表达依次为空白对照组的1.46倍、1.53倍。中药低、高剂量组均可显著抑制BALB/C裸鼠H1650肺癌种植瘤瘤体生长(P<0.05),抑瘤率分别为19.54%、47.03%。中药高、低剂量组CHOP/GADD153蛋白表达分别为模型组的1.59倍、1.44倍(P<0.05),GADD34蛋白表达量分别为模型组的1.98倍、1.49倍(P<0.05)。【结论】益气除痰方可抑制肺腺癌H1650种植瘤瘤体增长,促进H1650肺腺癌细胞凋亡,其机制可能与其能调控CHOP/GADD153、GADD34在内的内质网应激反应相关。

Objective To study the anti-tumor effect and mechanism of Yiqi Chutan Recipe （YCR）, a Chinese herbal prescription has the actions of nourishing qi and eliminating phlegm, on H1650 lung cancer. Methods The apoptotic rate of H1650 lung cancer cells before and after treatment with serum containing YCR was detected by flow cytometry. Western blotting method was used to detect the protein expression of growth arrest and DNA-damage inducible gene 153 （CHOP/GADD153） and GADD34 in H1650 cells of each group. BALB/C nude mice were used for the establishment of H1650 lung cancer xenografts model. The model nude mice were randomly divided into model group, low-dose YCR group （9.1 g·kg^-1·d^-1） and high-dose YCR group （27.3 g·kg^-1·d^-1） . The tumor size and mass were measured in each group, and the protein expression of CHOP/ GADD153 and GADD34 in H1650 tumor tissues of nude mice was detected by Western blotting method. Results The results of cell model test showed that apoptotic rate of H1650 cells was （8.14 ± 1.10） % in 15% （volume fraction） YCR-serum group and was （18.21 ± 3.07） % in 30% YCR-serum group. The expression of CHOP/ GADD153, GADD34 in 30% YCR-serum group was 1.46, 1.53 times as much as that of the blank control group. In in-vivo test, low- and high-dose YCR markedly inhibited the growth of H1650 lung tumor in BALB/ C nude mice （P〈0.05）, with the tumor-inhibition rate being 19.54%, 47.03% respectively. The expression level of CHOP/GADD153 in high-dose YCR group and low-dose YCR group was 1.59, 1.44 times as much as that of the model group （P〈0.05）, and the expression level of GADD34 was 1.98, 1.49 times as much as that of the model group （P〈0.05）. Conclusion YCR can inhibit the growth of H1650 lung cancer and promote the apoptosis of transplanted H1650 lung cancer cells, and its mechanism may be related to the regulation of endoplasmic reticulum stress response through the up-regulation of CHOP/GADD153 and GADD34 protein expression.