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Application of VGLL4 gene in treatment of tumors and related medicament of VGLL4 gene

A gene and drug technology, applied in the biological field, can solve problems such as excessive cell proliferation and increased risk of cancer

Active Publication Date: 2015-04-29
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the disorder of Hippo signaling pathway can cause anti-apoptotic effect, make cells hyperproliferate, and increase the risk of cancer

Method used

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  • Application of VGLL4 gene in treatment of tumors and related medicament of VGLL4 gene
  • Application of VGLL4 gene in treatment of tumors and related medicament of VGLL4 gene
  • Application of VGLL4 gene in treatment of tumors and related medicament of VGLL4 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1 The relationship between the expression of VGLL4 and the occurrence and development of gastric cancer

[0092] 1. Experimental method

[0093] In order to study the clinical relationship between VGLL4 and YAP, the mRNA levels of VGLL4 and YAP were detected in 84 cases of human gastric cancer samples and 91 cases of normal human gastric tissue samples. The total RNA was extracted with guanidine isothiocyanate in one step, the first strand was synthesized by AMV reverse transcriptase, and PCR amplification was performed with the corresponding primers under the action of DNA taq enzyme using AMV reverse transcriptase; see Table 1 for the PCR primers, Realtime - See Table 2 for the PCR reaction system, see Table 3 for the Realtime-PCR amplification program, and see Table 3 for the PCR amplification results figure 1 A and figure 1 b.

[0094] Table 1 Primer Sequence

[0095]

[0096]Table 2 Realtime-PCR reaction system

[0097]

[0098] Table 3 PCR react...

Embodiment 2

[0106] Example 2 VGLL4 inhibits YAP-mediated TEAD4 transcriptional activation and cell proliferation

[0107] 1. Purpose of the experiment

[0108] The purpose of this example is to study the regulation mode among VGLL4, YAP and TEAD4 and their influence on cell proliferation.

[0109] 2. Experimental method

[0110] 2.1 Plasmid construction

[0111] 1) Construction of pCDH-YAP plasmid and pCDH-VGLL4 plasmid: One-step extraction of total RNA from human HEK293 cells with guanidine isothiocyanate, synthesis of the first strand by AMV reverse transcriptase, and using it as a template for high-fidelity DNA polymerization in Takara Enzyme PrimeSTAR PCR amplification was performed with the corresponding primers under the action of the corresponding primers; the PCR primers are shown in Table 4, the PCR reaction system is shown in Table 5, and the PCR amplification program is shown in Table 6.

[0112] Table 4 Primer sequences

[0113]

[0114] Table 5 PCR reaction system

...

Embodiment 3

[0137]Example 3 Vgl4-TEAD4 interaction affects Vgl4 inhibition of YAP-mediated transcriptional activation of TEAD4 and its downstream genes

[0138] 1. Experimental materials

[0139] 1.1 Construction of plasmid

[0140] 1) HT-pET-28a vector: commercialized pET-28a (Novagen) was used as a template, and the following primers were used for construction. The PCR amplification program is: 98°C 2min; PCR cycle: 98°C 10s, 58°C 10s, 68°C 1kb / min, 18 cycles; 68°C 10min. Use DpnI demethylase to remove the unmutated wild-type pET28a template, transform the PCR amplified fragment into DH5α competent cells, and use the characteristic of E. coli to repair the loss of the vector to circularize the entire fragment to form a vector. After screening, the HT-pET-28a vector with the same backbone as the original pET28a vector but with TEV restriction sites and common multiple cloning sites was obtained.

[0141] HT-s:

[0142] 5'-Gaaaacctgtattttcagggcgccatggatccggaattcaaaggcctacgtcgacgagctca...

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Abstract

The invention relates to the field of biotechnology, and in particular relates to an application of a VGLL4 gene in treatment of tumors and a related medicament of the VGLL4 gene. The invention discloses a separated VGLL4 gene and a homologous sequence thereof, a VGLL4 protein and an application of related protein of the VGLL4 protein in preparation or screening of tumor treatment medicaments. Researches discover that VGLL4 can inhibit the transcription of a TEAD4 target gene CDX2 highly expressed in stomach cancer by inhibiting YAP-mediated TEAD4 transcriptional activation, thereby achieving the aim of inhibiting proliferation of cancer cells at last. The VGLL4 gene provided by the invention can be used for preparing medicaments for inhibiting the growth of tumor cells, developing anti-cancer medicaments and preparing medicaments for early diagnosis of cancer and medicaments for operation prognosis, so that the VGLL4 gene provides a new technical means for treating cancers, and has a great market value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of VGLL4 gene in treating tumors and related medicines. Background technique [0002] Gastric cancer is the second most common cancer in the world after lung cancer, with poor prognosis and high mortality. However, the molecular mechanism of gastric cancer is still unclear, and there is no effective detection method to detect early gastric cancer. The proto-oncogenes and tumor suppressor genes related to gastric cancer still need to be further studied. [0003] In recent years, studies have found that the key transcriptional coactivator YAP in the Hippo signaling pathway is significantly upregulated in lung cancer, liver cancer, colon cancer, pancreatic cancer, and breast cancer, and is a proto-oncogene. [0004] The Hippo signaling pathway is involved in physiological processes such as cell proliferation, cell differentiation, and apoptosis, and plays a vital role in the re...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/19C07K14/715C07K7/06C12Q1/68A61P35/00
Inventor 周兆才张雷季红斌赵允
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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