Journal of Jilin University(Medicine Edition) ›› 2021, Vol. 47 ›› Issue (4): 999-1007.doi: 10.13481/j.1671-587X.20210425

• Research in clinical medicine • Previous Articles     Next Articles

Establishment of human serous ovarian cancer cell culture system in vitro and its application in chemotherapeutic drug sensitivity detection

Jingjing GE1,Hongxia XU1,2,Lixia XIE1,Kaili DU1,Ming SANG1,2,3(),Xiaodong SUN1,2,3()   

  1. 1.Center for Translational Medicine,Xiangyang No. 1 People’s Hospital,Hubei University of Medicine,Xiangyang 441000,China
    2.Hubei Clinical Research Center of Parkinson’s Disease,Xiangyang No. 1 People’s Hospital,Hubei University of Medicine,Xiangyang 441000,China
    3.Hubei Key Laboratory of Wudang Local Chinese Medicine Research,Hubei University of Medicine,Shiyan 442000,China
  • Received:2020-11-30 Online:2021-07-28 Published:2021-07-22
  • Contact: Ming SANG,Xiaodong SUN E-mail:smxd2000@126.com;sunxiaodongsm@126.com

Abstract: Objective

To establish a culture system in vitro of human serous ovarian cancer cells and to theoretical basis to provide the experiment model for the clinical adminstration of the patients with serous ovarian cancer.

Methods

The human serous ovarian cancer cells were isolated and cultured from tumor tissue by type Ⅱ collagenase digestion and differential centrifugation method,and the third of generation serous ovarian cancer cells were used in subsequent experiments. The morphology of ovarian cancer cells was observed under inverted phase contrast microscope, and the cell growth curve was plotted and analyzed. The expression amounts of the human cancer antigen 125 (CA125), paired box gene 8 (PAX8) and epididymis protein 4 (HE4) in the cells, and the expression amounts of embryonic stem cell homologous transcription factor (Nanog) and octamer binding transcription factor 4 (Oct-4) in the cancer stem cells were detected by immunofluorescence. The drug sensitivity test was divided into 5-fluorouracil (5-FU) groups (0, 0.625, 1.250, 2.500, 5.000 mg·L-1), paclitaxel (PTX) groups (0, 0.062 5, 0.125 0, 0.250 0, 0.500 0 mg·L-1), temozolomide (TMZ) groups (0, 0.125, 0.250, 0.500, 1.000 mg·L-1), carboplatin (CBP) groups (0, 0.625, 1.250, 2.500, 5.000 mg·L-1), adriamycin (DOX) groups (0, 0.125, 0.250, 0.500, 1.000 mg·L-1) and melatonin (MLT) groups (0, 0.625, 1.250, 2.500, 5.000 mg·L-1);among them,0 mg·L-1 groups were used as control group; CCK-8 method was used to evaluate the survival rates of the cells.

Results

The primary serous ovarian cancer cells began to adhere to bottom of culture bottles 4 h after enzymatic digestion. The cells grew rapidly and had uniform morphology during continuous passages. The cells grew in a linearly phase during 2-7 d after inoculation, and grow and fuse into pieces after 8-10 days of culture. The results of immunofluorescence assay showed that CA125, PAX8,HE4, Nanog and Oct-4 were highly expressed in the cells. The drug sensitivity test showed that the ovarian cancer cells had high sensitivity to 5-FU,TMZ,CBP and MLT; compared with corresponding control groups,the survival rates of cells in different concentrations of 5-FU groups,TMZ groups and MLT groups and 0.0625 and 0.125 mg·L-1 PTX groups were significantly decreased(P<0.05 or P<0.01).

Conclusion

The culture system in vitro of human serous ovarian cancer cells is successfully established.The cells are sensitive to 5-FU, TMZ, CBP and MLT, which is of guiding significance for the selection of postoperative chemotherapy drugs in the patients with serous ovarian cancer.

Key words: ovarian neoplasms, primary cell culture, chemotherapeutic drug sensitivity, tumor markers, tumor stem cells

CLC Number: 

  • R737.31