Application of RFPL1S-201 in preparation of drugs for inhibiting proliferation, invasion and/or metastasis of ovarian cancer

A technology of RFPL1S-201, 1.RFPL1S-201, applied in the direction of drug combination, anti-tumor drugs, pharmaceutical formulations, etc.

Active Publication Date: 2020-09-04
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although without protein-coding potential, lncRNAs have been

Method used

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  • Application of RFPL1S-201 in preparation of drugs for inhibiting proliferation, invasion and/or metastasis of ovarian cancer
  • Application of RFPL1S-201 in preparation of drugs for inhibiting proliferation, invasion and/or metastasis of ovarian cancer
  • Application of RFPL1S-201 in preparation of drugs for inhibiting proliferation, invasion and/or metastasis of ovarian cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Cell lines

[0045] The ovarian cancer cell line SKOV3 was purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences; the normal ovarian epithelial cell line IOSE386, and the ovarian cancer cell lines A2780, HO-8910, HO-8910PM, OVCAR3, and OVCAR8 were purchased from KGI. Cell culture conditions are as follows:

[0046] Culture medium: SKOV3: McCoy’s 5A incomplete medium (1×) + 10% fetal bovine serum (FBS)

[0047] A2780: DMEM incomplete high-glucose medium (1×) + 10% fetal bovine serum (FBS)

[0048] Digestive solution: 0.25% Trypsin-EDTA (1×)

[0049] Freezing medium: fetal bovine serum (FBS): dimethyl sulfoxide (DMSO): = 9: 1

[0050] 2. Clinical specimens

[0051] The samples in this study were collected from patients undergoing surgery in Nanjing Maternal and Child Health Hospital from 2016 to 2017. A total of 13 normal ovarian tissue samples (from cervical cancer and endometrial cancer with preventive ovary rese...

Embodiment 2

[0061] 1. Construction of RFPL1S-201 interference RNA and overexpression plasmid:

[0062] The interfering RNA sequence of RFPL1S-201 and the control interfering RNA sequence were designed and synthesized by American Thermo Fisher Company.

[0063] The RFPL1S-201 overexpression plasmid pc-RFPL1S-201 was constructed and synthesized by Shanghai Jierui Bioengineering Co., Ltd. on the basis of pcDNA3.1(+), the cloning site is: NheI / NotI, and LncRNA RFPL1S-201 (ENST00000419368. 1) The full length is linked to the cytomegalovirus (CMV) promoter of the pcDNA3.1 vector. Use the OMEGA kit (D6915-03) plasmid to measure the concentration of the extracted plasmid, and detect that the sequence is connected successfully, and use it directly for subsequent experiments or store it at -80°C.

[0064] 2. Cell Transfection

[0065] 1) When the confluence of SKOV3 and A2780 cells cultured in the 6-well plate reached 90%-95%, the RFPL1S-201 overexpression plasmid, control pcDNA3.1 plasmid, RFPL1...

Embodiment 3

[0072] Example 3 Cell Proliferation Experiment

[0073] 1) Count the cells after 24 hours of transient transfection, dilute to the desired concentration with medium containing 10% FBS, and then add the cell suspension to a 96-well plate. Each well contains about 1000 cells in 100μ suspension, and divides into 0 hours. , 24h, 48h, 72h four groups, 5-6 auxiliary wells for each group, placed at 37°C, 5% CO 2 Wait for the cells to adhere to the wall for 3-4 hours in the incubator;

[0074] 2) Measure the cell viability according to the grouping time after the cells adhere to the wall: replace the medium with a mixed solution with 10% CCK-8 reagent, 37°C, 5% CO 2 After reacting in the incubator for 1 hour, detect on a microplate reader, detection wavelength: 450nm.

[0075] see results figure 2 A-B and image 3 A-B; It can be seen from the figure that overexpression of RFPL1S-201 can significantly inhibit the proliferation of ovarian cancer cells, and knockdown of RFPL1S-201 c...

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Abstract

The invention discloses application of RFPL1S-201 in the preparation of drugs for inhibiting the proliferation, invasion and/or metastasis of ovarian cancer, application of RFPL1S-201 or substances that promote the expression of RFPL1S-201 in the preparation of drugs for inhibiting the proliferation, invasion and/or metastasis of ovarian cancer, and application of expression plasmids overexpressing RFPL1S-201 in the preparation of drugs for inhibiting the proliferation, invasion and/or metastasis of ovarian cancer. It is found that lncRNA RFPL1S-201 can inhibit the proliferation, invasion andmetastasis of ovarian cancer cells, and meanwhile lncRNA RFPL1S-201 can inhibit the liver metastasis of ovarian cancer in vivo. A mechanism of action is mainly to inhibit tumor progression and promotetumor chemotherapy sensitivity by inhibiting an IFN-STAT1 signal channel.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of RFPL1S-201 in the preparation of drugs for inhibiting proliferation, invasion and / or metastasis of ovarian cancer. Background technique [0002] Ovarian cancer is a common malignant tumor of the female reproductive system, which can occur at any age. Due to atypical early symptoms, rapid progression, and early metastasis of pelvic implants, the mortality rate has always been the highest among gynecological tumors, seriously threatening women's lives[1]. Cytoreductive surgery combined with platinum-based chemotherapy is the main clinical treatment. With the improvement of surgical skills and chemotherapy, the short-term survival rate of ovarian cancer patients has increased significantly. However, the long-term recurrence rate is high and the prognosis is still poor. . Ovarian cancer's extremely high metastasizing ability and extremely high drug resistance are one of th...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/713A61P35/00A61P35/04
CPCA61K31/713A61K45/00A61P35/00A61P35/04
Inventor 贾雪梅徐娟刘丝雨许鹏飞葛莉莉黄可
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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