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Application of KCP gene in cervical cancer taxol sensibility detection

A paclitaxel, sensitive technology, applied in the field of biomedicine, can solve the problems of treatment failure, insensitivity, disease progression, etc., and achieve the effect of improving the effect of chemotherapy

Active Publication Date: 2019-07-09
BEIJING OBSTETRICS & GYNECOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies in the United States and Japan have reported that 20% of patients with locally advanced cervical cancer are insensitive to paclitaxel-based neoadjuvant chemotherapy, leading to treatment failure
[0004] At present, there is no specific marker for evaluating paclitaxel sensitivity. If experimental chemotherapy is performed for paclitaxel-insensitive cervical cancer patients, it will increase the patient's economic costs and side effects of chemotherapy, and even lead to disease progression

Method used

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  • Application of KCP gene in cervical cancer taxol sensibility detection
  • Application of KCP gene in cervical cancer taxol sensibility detection
  • Application of KCP gene in cervical cancer taxol sensibility detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Genome-wide CRISPR / Cas9 knockout technology screening paclitaxel chemosensitivity genes in SiHa cells

[0025] The genome-wide CRISPR / Cas9 library lentivirus infected cervical cancer SiHa cells, co-cultured with paclitaxel chemotherapy drug (drug group) and DMSO solvent (control group) for 7 days and 14 days respectively, high-throughput sequencing analysis results (RNA of KCP transcripts The sequences are respectively SEQ ID NO:2 (the first), SEQ ID NO:3 (the second) and SEQ ID NO:4 (the third)), select the fold change (Fold Change, FC), Bayesian Factor (Bayes Factor, BF) Genes that meet the following screening conditions are regarded as candidate genes.

[0026] Filter criteria:

[0027] In drug group FC Paclitaxel ≤ -2, in control |FC DMSO |<2; and

[0028] in drug group BF Paclitaxel >2, BF in control group DMSO <0.

[0029] The genes that meet the above screening conditions indicate that the knockout of this gene leads to an increase in the sensiti...

Embodiment 2

[0032] Example 2 High Content Screening (High Content Screening, HCS) (mixed target, single target) increases paclitaxel chemosensitivity genes after knockout

[0033]According to the results of HCS mix, 42 sgRNA targets (6 targets per gene) of 7 positive genes were selected, plasmids were extracted, packaged into lentiviruses, infected cells, and entered into the HCS single-target screening experiment, 4ng / Compared with the control group (without paclitaxel) treated with mL paclitaxel, the proliferation results of each group were analyzed by Celigo cell counting as follows (see Table 1).

[0034] Table 1. High-content screening (single target) paclitaxel-related genes

[0035]

[0036]

[0037] Note: with the suffix "-4ng" means adding 4ng / ml paclitaxel drug, without the suffix means the control group without paclitaxel.

[0038] The results of this round of high-content screening showed that the cells in the NC group (SiHa) proliferated normally, and the proliferatio...

Embodiment 3

[0039] Example 3 Study on the function of KCP gene on paclitaxel-induced cervical cancer cells (for the above KCP single target (05003-1))

[0040] Normal target cells, as well as cells infected with sgRNA negative control virus and sgRNA virus with KCP gene, were treated with paclitaxel ("+" indicates the addition of paclitaxel). After adding paclitaxel for 1-5 days, the MTT test results showed that compared with the sgCtrl+ / sgCtrl group, the cells in the sgKCP+ / sgKCP group were more sensitive to paclitaxel, and their proliferation was significantly inhibited (P diagram 2-1 ), the cell cloning ability was significantly weakened (P Figure 2-2 ), but there was no significant difference in the apoptosis rate (P>0.05) (see Figure 2-3 ).

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Abstract

The invention relates to application of a KCP gene in preparation of a cervical cancer taxol treatmetn sensibility detection product. The detection product is a detection reagent or a detection kit. The detection reagent comprises a KCP gene sequence integrity detection reagent and / or a KCP gene expression product detection reagent. The detection product evaluates the sensitivity of a tested cervical cancer patient to taxol drug chemotherapy by detecting the sequence integrity of the KCP gene or the expression product of the KCP gene, a cervical cancer treatment new target spot is provided, and the application assists in the effect of taxol chemotherapy on the cervical cancer patient in advance.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of KCP gene. Background technique [0002] Cervical cancer is the fourth most common malignant tumor that threatens women's health worldwide. According to statistics in 2012, there are 530,000 new cases of cervical cancer and 270,000 new deaths worldwide each year. my country is the country with the largest number of new cases and deaths, with 131,500 new cases and 53,000 new deaths each year, accounting for 18.4% of the female malignant tumor deaths in my country. In underdeveloped countries, the 5-year survival rate of cervical cancer is not high <50%, especially in stage III-IV, the survival rate is only 5%-15%. Cervical cancer seriously threatens women's lives. Although there has been a perfect cervical cancer screening system for many years, early detection and early treatment can be achieved. However, based on my country's national conditions, nearly half of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/106C12Q2600/158C12Q2600/156
Inventor 吴玉梅何玥
Owner BEIJING OBSTETRICS & GYNECOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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